Nattokinase is a potent fibrinolytic enzyme with the potential for fighting cardiovascular diseases. In this study, Bacillus subtilis natto were immobilized in the alginate – chitosan complex for fermentation of nattokinase enzyme. Six factors affecting the efficiency of immobilization cells were screened by Plackett – Burman design including: concentration of alginate, concentration of chitosan, pH of chitosan, concentration of CaCl2, added cells density, shaking time after supplementing chitosan. Results of optimization have identified two factors affecting the efficiency of cell immobilization. They are concentration of alginate (2.5%) and added cells density (approximately 5.86 million colonies per milliliter). With these two factors optimized and others kept at the normal level, immobilization efficiency reached 90.73%. After Bacillus subtilis natto had been immobilized by optimization of parameters, we conducted application for fermenting nattokinase. For 24 hours of fermentation, nattokinase enzyme activity reached 71.80 ± 0.19 FU/ml. Immobilized Bacillus subtilis natto cells were reused 6 times and on the 6 th time of reuse, nattokinase enzyme activity only decreased 2.7% in compared with the 1st reuse.
Optimization Of The Possibility Synthetic Nattokinase In Soybean Substrates T...inventionjournals
Nattokinase is an enzyme with strong fibrinolytic activity that can be used for preventing thrombolytic diseases. In this study, we make survey the effect fermentation conditions to B.subtilis natto strain on the soybean substrate to enhance the nattokinase activity and optimization biosynthesis capabilities of nattokinase by Plackett-Burman experimental combined with response surface methodology RSM-CCD. The optimal results received nattokinase activity 136.6 FU/g. We also try to create dried products by freeze drying process in the conditions -80°C, 24 hours, 6-7 Pa. After that we examined moisture and nattokinase activity in these product. The results of experiments show that moisture was 4.3%, nattokinase activity was 515 FU/g. This research help expand application for creating soybeans powder products with hight nattokinase activity.
IJRET : International Journal of Research in Engineering and Technology is an international peer reviewed, online journal published by eSAT Publishing House for the enhancement of research in various disciplines of Engineering and Technology. The aim and scope of the journal is to provide an academic medium and an important reference for the advancement and dissemination of research results that support high-level learning, teaching and research in the fields of Engineering and Technology. We bring together Scientists, Academician, Field Engineers, Scholars and Students of related fields of Engineering and Technology
Phytochemical analysis, protein content & antimicrobial activities of sel...eSAT Journals
Abstract Two seed samples of Glycine max Linn. (S1, S2) were purchased from two retail stores of local market. Non-sprouted and sprouted seed powder were extracted separately with methanol (100%, 50%) by cold maceration to obtain methanolic and hydroalcoholic extract of Glycine max Sample 1 was designated as MES1 and HES1 and sample 2 as MES2 and HES2 respectively. Phytochemical analysis indicated the presence of various phytoconstituents viz. phytosterols, flavonoids, phenolic compounds, tannins, carbohydrates, proteins, amino acids, fixed oils and fats etc. Thin layer chromatography study on extracts revealed the presence of a number of compounds. The protein content of these samples were studied. The protein content of samples MES1, HES1, MES2 and HES2 with respect to BSA was found to be 90.6 2μg/ml, 82μg/ml, 94.5μg/ml and 79.1μg/ml respectively. The highest among these were found to be in MES2. Sprouting enhanced the protein content of the two samples. The samples have shown antimicrobial activity at selected concentration and microbial strains (26mm) for gram negative bacteria (27mm) for gram positive bacteria. Keywords: Glycine max Linn, phytochemical constituents, TLC, antimicrobial activity, protein, methanolic extract, hydroalcoholic extract.
Royal jelly accelerates recovery from oral mucositisBee Healthy Farms
Great news for cancer patients who suffer from oral mucositis. However, individuals undergoing this type of intense therapy also need to consume large quantities of propolis to protect their weakened immune system.
Optimization Of The Possibility Synthetic Nattokinase In Soybean Substrates T...inventionjournals
Nattokinase is an enzyme with strong fibrinolytic activity that can be used for preventing thrombolytic diseases. In this study, we make survey the effect fermentation conditions to B.subtilis natto strain on the soybean substrate to enhance the nattokinase activity and optimization biosynthesis capabilities of nattokinase by Plackett-Burman experimental combined with response surface methodology RSM-CCD. The optimal results received nattokinase activity 136.6 FU/g. We also try to create dried products by freeze drying process in the conditions -80°C, 24 hours, 6-7 Pa. After that we examined moisture and nattokinase activity in these product. The results of experiments show that moisture was 4.3%, nattokinase activity was 515 FU/g. This research help expand application for creating soybeans powder products with hight nattokinase activity.
IJRET : International Journal of Research in Engineering and Technology is an international peer reviewed, online journal published by eSAT Publishing House for the enhancement of research in various disciplines of Engineering and Technology. The aim and scope of the journal is to provide an academic medium and an important reference for the advancement and dissemination of research results that support high-level learning, teaching and research in the fields of Engineering and Technology. We bring together Scientists, Academician, Field Engineers, Scholars and Students of related fields of Engineering and Technology
Phytochemical analysis, protein content & antimicrobial activities of sel...eSAT Journals
Abstract Two seed samples of Glycine max Linn. (S1, S2) were purchased from two retail stores of local market. Non-sprouted and sprouted seed powder were extracted separately with methanol (100%, 50%) by cold maceration to obtain methanolic and hydroalcoholic extract of Glycine max Sample 1 was designated as MES1 and HES1 and sample 2 as MES2 and HES2 respectively. Phytochemical analysis indicated the presence of various phytoconstituents viz. phytosterols, flavonoids, phenolic compounds, tannins, carbohydrates, proteins, amino acids, fixed oils and fats etc. Thin layer chromatography study on extracts revealed the presence of a number of compounds. The protein content of these samples were studied. The protein content of samples MES1, HES1, MES2 and HES2 with respect to BSA was found to be 90.6 2μg/ml, 82μg/ml, 94.5μg/ml and 79.1μg/ml respectively. The highest among these were found to be in MES2. Sprouting enhanced the protein content of the two samples. The samples have shown antimicrobial activity at selected concentration and microbial strains (26mm) for gram negative bacteria (27mm) for gram positive bacteria. Keywords: Glycine max Linn, phytochemical constituents, TLC, antimicrobial activity, protein, methanolic extract, hydroalcoholic extract.
Royal jelly accelerates recovery from oral mucositisBee Healthy Farms
Great news for cancer patients who suffer from oral mucositis. However, individuals undergoing this type of intense therapy also need to consume large quantities of propolis to protect their weakened immune system.
IJRET : International Journal of Research in Engineering and Technology is an international peer reviewed, online journal published by eSAT Publishing House for the enhancement of research in various disciplines of Engineering and Technology. The aim and scope of the journal is to provide an academic medium and an important reference for the advancement and dissemination of research results that support high-level learning, teaching and research in the fields of Engineering and Technology. We bring together Scientists, Academician, Field Engineers, Scholars and Students of related fields of Engineering and Technology.
(48)Screening and identifiation of Bacillus sp. isolated from traditional Vie...minhdaovan
Fibrinolytic enzymes produced by microorganisms have been attractive in prevention and
treatment of cardiovascular diseases (CVDs) by their low-cost and safety. This study focused
on screening for the existence of firinolytic enzymes in Vietnamese traditional fermented
soybean paste products and isolation and identifiation of related bacteria. Sixteen fermented
soybean paste samples were collected over three regions of Vietnam in which seven samples
gave the positive results on firinolytic enzyme activity. Miso (MS) and Green Chili (GC)
samples had the highest firinolytic enzyme activities (1.81 and 0.77 FU/g, respectively).
According to morphological features, four strains of bacteria were isolated and all of them
were found to produce firinolytic enzymes. The enzyme activities produced by four isolated
strains were in a range of 29.7 - 77.9 FU/g after culturing on solid state media for 24 h. The
isolated strains were identifid as Bacillus amyloliquefaciens using 16 rRNA sequence and
phylogenetic analysis with 99% similarity
ABSTRACT- Aberrant glycosylation has been recognized as hallmark of cancer. Exploiting differences in glycosylation between malignant and healthy tissues offers excellent opportunities to identify sensitive and specific cancer biomarkers. Plant lectins have demonstrated the ability to specifically agglutinate malignant transformed cells. Lectins are sugar binding proteins or glycoprotein of non-immune origin which agglutinate cells or precipitate glycol-conjugates. Some lectins shown to the anti- proliferative effect on cancer cells. A wide scope of this application of lectins is that it can be used for diagnosis as well as therapeutics of cancer. The objective of the present study was to purify a lectin from tubers of Arisaema intermedium and evaluate in vitro anti-proliferative potential towards HCT-15, a human colon cancer cell line. The present study was conceived as an offshoot to the ongoing work on lectins in our laboratory. The already reported Arisaema intermedium (AIL) lectin was purified on asialofetuin linked amino-activated silica bead matrix. The purity of the affinity purified lectin was ascertained by SDS-PAGE, pH-8.3. The lectin activity was assessed by hemagglutination and protein concentration was determined by Lowry’s method. The cytotoxicity of AIL towards HCT-15 was evaluated by MTT assay. The mechanism of anti-proliferative effect was assessed by evaluation of cell morphology, trypan blue exclusion assay, DNA fragmentation and nucleic acid content determination.
Key-words- Araceae, Arisaema, Asialofetuin, Antiproliferative effect, Apoptosis, Cytotoxicity Lectins, Mechanistic
Bioconversion of Penicillin to CephalosporinIOSR Journals
Cephalosporins are known as 3rd generation broad spectrum Beta lactam antibiotics, which can also be produced synthetically. Commonly, chemical ring expansion followed by an enzymatic removal of the phenylacetyl side chain is commonly employed to convert penicillin G into 7-aminodeacetoxycephalosporanic acid, the precursor for the manufacture of semisynthetic cephalosporins. This process requires several steps, is expensive and highly polluting. Thus there is a need to device a simple biological route to replace the chemical process. A mutant of Streptomyces clavuligerus NP1 was reported to converts Penicillin G to Deacetoxycephalosporin G (DAOG;phenylacetyl-7-aminodeacetoxycephalosporanic acid) enzymatically[5,8] . This enzyme, deacetoxycephalosporin synthase has the potential for the large scale transformation of Penicillin G to deacetoxycephalosporin. The present work studies the conditions required for efficient transformation of Penicillin G to Deacetoxycephalosporin using the wild type strain Streptomyces clavuligerus . Detection of cephalosporin was carried out using various methods. Additionally succinic acid formation was also studied as it could be used as a commercially important by product of the transformation. Deacetoxycephalosporin synthase also extracted and partially purified and characterised.
Effcet of Vernonia Amygdalina.Del Ethanolic Extract Fraction on Serum Prolact...iosrjce
IOSR Journal of Agriculture and Veterinary Science (IOSR-JAVS) is a double blind peer reviewed International Journal edited by the International Organization of Scientific Research (IOSR). The journal provides a common forum where all aspects of Agricultural and Veterinary Sciences are presented. The journal invites original papers, review articles, technical reports and short communications containing new insight into any aspect Agricultural and Veterinary Sciences that are not published or not being considered for publication elsewhere.
Enzymatic Saccharification of Lignocellulosic BiomassBiorefineryEPC™
Enzymatic Saccharification of Lignocellulosic Biomass
YOU AGREE TO INDEMNIFY BiorefineryEPCTM , AND ITS AFFILIATES, OFFICERS, AGENTS, AND EMPLOYEES AGAINST ANY CLAIM OR DEMAND, INCLUDING REASONABLE ATTORNEYS' FEES, RELATED TO YOUR USE, RELIANCE, OR ADOPTION OF THE DATA FOR ANY PURPOSE WHATSOEVER. THE DATA ARE PROVIDED BY BiorefineryEPCTM "AS IS" AND ANY EXPRESS OR IMPLIED WARRANTIES, INCLUDING BUT NOT LIMITED TO THE IMPLIED WARRANTIES OF MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE ARE EXPRESSLY DISCLAIMED. IN NO EVENT SHALL BiorefineryEPCTM BE LIABLE FOR ANY SPECIAL, INDIRECT OR CONSEQUENTIAL DAMAGES OR ANY DAMAGES WHATSOEVER, INCLUDING BUT NOT LIMITED TO CLAIMS ASSOCIATED WITH THE LOSS OF DATA OR PROFITS, WHICH MAY RESULT FROM ANY ACTION IN CONTRACT, NEGLIGENCE OR OTHER TORTIOUS CLAIM THAT ARISES OUT OF OR IN CONNECTION WITH THE USE OR PERFORMANCE OF THE DATA.
1. bioconversion of progesterone using aqueous two phase system by comamonas ...Darshan Rudakiya
Strains of Comamonas acidovoras have been
reported to have vital role in degradation of natural
as well as complex organic compounds.
Comamonas acidovoras MTCC 3364 has been
routinely reported for steroid bioconversion by
activated beads in aqueous system. Previously
studies observed that progesterone was converted
into AD and ADD steroids. These compounds were
higher valuable steroids which were mainly used in
different types of drugs. Novel system was used for
the bioconversion of progesterone in to AD and
ADD. Aqueous two-phase system was initially
optimized for the progesterone solubility, separation
of two phases and extraction of product steroids
from the system. Even 1% of PEG can easily
dissolved progesterone but 5% of PEG needed for
separating phases and for extraction purposes.
Beads was easily tolerate up to 20% of PEG system
even for 120 hours. Hexane used as better
extraction solvent among different type of solvent
system. Products were shown in PEG 6000 system
after 96 hours in fewer amounts. PEG 8000 system
showed products 48 hours in minor amount but
increased in 96 hours. Advantage of aqueous twophase
system in progesterone bioconversion was
better stability of progesterone, extraction of steroid
products and stability of the system.
Optimizing some conditions for spray drying in synbiotic capsule from Bacillu...inventionjournals
The study is of determination of various conditions for spray drying in producing synbiotic in the form of capsule from Bacillus subtilis natto. The experiments were conducted to examine effects of various factors such as the resistant starch-to-matodextrin ratio before drying, the inlet gas temperature and the inlet flow. The optimization experiments are administered: a ratio of wall materials to the core material is 5% (w/v) in which the ratio of resistant starch and maltodextrin is 1:9, the inlet gas temperatureis 1100C, the inlet flow is 5.60 ml/minute and the spray pressure is 2 bar. In the condition for spray drying as mentioned earlier, the initial step for trial production of synbiotic capsules was conducted with the cell density of B.subtilis natto being 8.55 ± 0.18 log(CFU/g), the activating effect of nattokinase is 518.2 FU/g and the moisture is 9.11%. During 60 days’ preservation of the product, the resulting indexes such as the cell density of B.subtilis natto in the capsule, the activating effect of nattokinase and the moisture are stable.
Validation Of Radiation Sterilization Dose For Proteases Immobilized On Aldeh...iosrphr_editor
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
Comparison of Three Slaughtering Methods of Goat on Carcass and Prime Cuts Re...inventionjournals
Procedures employed on slaughtering of goat varies due to lack of information on the effect of each process such as scalding, flaying and singeing on the different slaughter parameters and prime cut recovery. Using fifteen male Anglo-nubian goats at 8months old, the effect on slaughtering weight, hot carcass, chilled carcass, dressing percentage and drip loss as well as in the prime cut production was documented. Results show that on slaughter weight, there is no significant difference obtained due to the uniform management production system used in raising the animals. On hot and chilled carcass weights, significant difference was found (p=0.01). Difference on dressing percentage revealed that singeing has the highest recovery rate at 59.92%, and values obtained is significantly different (p=0.05). Drip loss is also highly significant (p=0.05). On the prime cut recovery, highly significant differences were obtained on percentage (%) shoulder, loin and flank, offals and the skin and other cuts did not vary significantly from each other. It can therefore be concluded that each of procedures employed has an effectto the differentparameters considered in goat slaughtering and in prime cut recovery.
Cell-free DNA Levels Serum Patients with Benign and Malignant Epithelial Ovar...inventionjournals
An elevated level of cell-free DNA (cfDNA) in the blood circulation has detected in cancer patients in comparison with healthy controls. CfDNA circulation in plasma and serum extensively studied and the results are highly variable due to many factors influence the test results that was preanalytic factors as well as analytic factors. Objectives: Is there any difference in the concentration of serum DNA among patients with benign epithelial ovarian tumors and malignant epithelial ovarian tumors? What is the clinicopathological variable that influences the cfDNA circulation? Method: Venous blood drawn with plain vacutainer, centrifuged at 1,000 rpm for 30 minutes, serum kept in -800 C freezer. The cfDNA extracted used NaI method.Results: Collected 30 cases of the benign ovarian tumor and 54 cases of malignant ovarian tumors. The average level serum cfDNA of benign epithelial ovarian tumors and malignant epithelial ovarian tumors were 24.6 ng/mL and 22:29 ng/mL respectively and statistically was not significantly different (p = 0.64). In multivariable analysis with linear regression, there were no clinicopathological variables that statistically significant influence the cfDNA levels in patients with epithelial ovarian tumors where p > 0.05. Conclusion: Concentration of cfDNA circulation of benign epithelial ovarian tumors a little bit higher than malignant epithelial ovarian tumors, but statistically was not significantly different. There was no clinicopathological variable influence the concentration of cfDNA circulation of ovarian tumors.
No, Orally Administered Hemolymph Of Limicolaria Aurora Does Not Reduce Blood...inventionjournals
The oral administration of the hemolymph of Limicolaria aurora at doses of 22.8 and 45.6 mg/kg body wt did not result in any significant difference in the systolic, diastolic, pulse pressure, mean arterial pressure and heart rate of both normotensive and adrenaline induced hypertensive wistar rats. The data from this study show the lack of antihypertensive potential associated with the oral ingestion of Limicolaria aurora hemolymph.
Assessment of Microbial Contamination of the Tooth Brush Head Used On Orthodo...inventionjournals
Introduction: Oral diseases can be greatly controlled by reducing the microbial load in the oral cavity and this can be achieved by maintaining proper oral hygiene.Tooth brushes are the most commonly used oral hygiene aid to promote oral health and prevent dental diseases. The insertion of fixed appliances alters the oral microbiological profile, thus increasing the risk for caries and gingivitis considerably. Aim: To assess the microbial growth of S.Mutans and Lactobacillus between and among the brushes. Setting and Study Design: A Hospital setting and Randomized Control study design Methods:A total of 56 (MB) patients aged 16-26 years received a toothbrush [Regular soft bristle design (group-A) and Orthodontic bristle design (group B)],A sterile gamma radiated pouch and checklist was distributed to each participant. After 2 weeks period the brushes were collected and placed in 5ml saline solution (0.05g Sodium Chloride). The suspension was incubated on selective agar plates and the amount of Streptococcus mutans and lactobacilli for each brush head was assessed. Results:The retention of S.Mutanswas found to be higher in group A, as compared to group B and was found to be statistically more significant between the two groups (P<0.001). The retention of Lactobacillus was also found to be higher in group A, as compared to group B and was found to be statistically significant between the groups (P= 0.001). However, there was no significant difference (P= 0.101) observedamong the microbial growth of S.Mutans and Lactobacillus in two bristle designs. Conclusions: Regular soft bristle design had a higher microbial load than those of subjects using orthodontic bristle design, a more frequent replacement of toothbrushes during t treatment may be advisable. Due to significant differences between the two bristle designs, the orthodontic toothbrush is recommended for patients undergoing orthodontic t appliances
IJRET : International Journal of Research in Engineering and Technology is an international peer reviewed, online journal published by eSAT Publishing House for the enhancement of research in various disciplines of Engineering and Technology. The aim and scope of the journal is to provide an academic medium and an important reference for the advancement and dissemination of research results that support high-level learning, teaching and research in the fields of Engineering and Technology. We bring together Scientists, Academician, Field Engineers, Scholars and Students of related fields of Engineering and Technology.
(48)Screening and identifiation of Bacillus sp. isolated from traditional Vie...minhdaovan
Fibrinolytic enzymes produced by microorganisms have been attractive in prevention and
treatment of cardiovascular diseases (CVDs) by their low-cost and safety. This study focused
on screening for the existence of firinolytic enzymes in Vietnamese traditional fermented
soybean paste products and isolation and identifiation of related bacteria. Sixteen fermented
soybean paste samples were collected over three regions of Vietnam in which seven samples
gave the positive results on firinolytic enzyme activity. Miso (MS) and Green Chili (GC)
samples had the highest firinolytic enzyme activities (1.81 and 0.77 FU/g, respectively).
According to morphological features, four strains of bacteria were isolated and all of them
were found to produce firinolytic enzymes. The enzyme activities produced by four isolated
strains were in a range of 29.7 - 77.9 FU/g after culturing on solid state media for 24 h. The
isolated strains were identifid as Bacillus amyloliquefaciens using 16 rRNA sequence and
phylogenetic analysis with 99% similarity
ABSTRACT- Aberrant glycosylation has been recognized as hallmark of cancer. Exploiting differences in glycosylation between malignant and healthy tissues offers excellent opportunities to identify sensitive and specific cancer biomarkers. Plant lectins have demonstrated the ability to specifically agglutinate malignant transformed cells. Lectins are sugar binding proteins or glycoprotein of non-immune origin which agglutinate cells or precipitate glycol-conjugates. Some lectins shown to the anti- proliferative effect on cancer cells. A wide scope of this application of lectins is that it can be used for diagnosis as well as therapeutics of cancer. The objective of the present study was to purify a lectin from tubers of Arisaema intermedium and evaluate in vitro anti-proliferative potential towards HCT-15, a human colon cancer cell line. The present study was conceived as an offshoot to the ongoing work on lectins in our laboratory. The already reported Arisaema intermedium (AIL) lectin was purified on asialofetuin linked amino-activated silica bead matrix. The purity of the affinity purified lectin was ascertained by SDS-PAGE, pH-8.3. The lectin activity was assessed by hemagglutination and protein concentration was determined by Lowry’s method. The cytotoxicity of AIL towards HCT-15 was evaluated by MTT assay. The mechanism of anti-proliferative effect was assessed by evaluation of cell morphology, trypan blue exclusion assay, DNA fragmentation and nucleic acid content determination.
Key-words- Araceae, Arisaema, Asialofetuin, Antiproliferative effect, Apoptosis, Cytotoxicity Lectins, Mechanistic
Bioconversion of Penicillin to CephalosporinIOSR Journals
Cephalosporins are known as 3rd generation broad spectrum Beta lactam antibiotics, which can also be produced synthetically. Commonly, chemical ring expansion followed by an enzymatic removal of the phenylacetyl side chain is commonly employed to convert penicillin G into 7-aminodeacetoxycephalosporanic acid, the precursor for the manufacture of semisynthetic cephalosporins. This process requires several steps, is expensive and highly polluting. Thus there is a need to device a simple biological route to replace the chemical process. A mutant of Streptomyces clavuligerus NP1 was reported to converts Penicillin G to Deacetoxycephalosporin G (DAOG;phenylacetyl-7-aminodeacetoxycephalosporanic acid) enzymatically[5,8] . This enzyme, deacetoxycephalosporin synthase has the potential for the large scale transformation of Penicillin G to deacetoxycephalosporin. The present work studies the conditions required for efficient transformation of Penicillin G to Deacetoxycephalosporin using the wild type strain Streptomyces clavuligerus . Detection of cephalosporin was carried out using various methods. Additionally succinic acid formation was also studied as it could be used as a commercially important by product of the transformation. Deacetoxycephalosporin synthase also extracted and partially purified and characterised.
Effcet of Vernonia Amygdalina.Del Ethanolic Extract Fraction on Serum Prolact...iosrjce
IOSR Journal of Agriculture and Veterinary Science (IOSR-JAVS) is a double blind peer reviewed International Journal edited by the International Organization of Scientific Research (IOSR). The journal provides a common forum where all aspects of Agricultural and Veterinary Sciences are presented. The journal invites original papers, review articles, technical reports and short communications containing new insight into any aspect Agricultural and Veterinary Sciences that are not published or not being considered for publication elsewhere.
Enzymatic Saccharification of Lignocellulosic BiomassBiorefineryEPC™
Enzymatic Saccharification of Lignocellulosic Biomass
YOU AGREE TO INDEMNIFY BiorefineryEPCTM , AND ITS AFFILIATES, OFFICERS, AGENTS, AND EMPLOYEES AGAINST ANY CLAIM OR DEMAND, INCLUDING REASONABLE ATTORNEYS' FEES, RELATED TO YOUR USE, RELIANCE, OR ADOPTION OF THE DATA FOR ANY PURPOSE WHATSOEVER. THE DATA ARE PROVIDED BY BiorefineryEPCTM "AS IS" AND ANY EXPRESS OR IMPLIED WARRANTIES, INCLUDING BUT NOT LIMITED TO THE IMPLIED WARRANTIES OF MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE ARE EXPRESSLY DISCLAIMED. IN NO EVENT SHALL BiorefineryEPCTM BE LIABLE FOR ANY SPECIAL, INDIRECT OR CONSEQUENTIAL DAMAGES OR ANY DAMAGES WHATSOEVER, INCLUDING BUT NOT LIMITED TO CLAIMS ASSOCIATED WITH THE LOSS OF DATA OR PROFITS, WHICH MAY RESULT FROM ANY ACTION IN CONTRACT, NEGLIGENCE OR OTHER TORTIOUS CLAIM THAT ARISES OUT OF OR IN CONNECTION WITH THE USE OR PERFORMANCE OF THE DATA.
1. bioconversion of progesterone using aqueous two phase system by comamonas ...Darshan Rudakiya
Strains of Comamonas acidovoras have been
reported to have vital role in degradation of natural
as well as complex organic compounds.
Comamonas acidovoras MTCC 3364 has been
routinely reported for steroid bioconversion by
activated beads in aqueous system. Previously
studies observed that progesterone was converted
into AD and ADD steroids. These compounds were
higher valuable steroids which were mainly used in
different types of drugs. Novel system was used for
the bioconversion of progesterone in to AD and
ADD. Aqueous two-phase system was initially
optimized for the progesterone solubility, separation
of two phases and extraction of product steroids
from the system. Even 1% of PEG can easily
dissolved progesterone but 5% of PEG needed for
separating phases and for extraction purposes.
Beads was easily tolerate up to 20% of PEG system
even for 120 hours. Hexane used as better
extraction solvent among different type of solvent
system. Products were shown in PEG 6000 system
after 96 hours in fewer amounts. PEG 8000 system
showed products 48 hours in minor amount but
increased in 96 hours. Advantage of aqueous twophase
system in progesterone bioconversion was
better stability of progesterone, extraction of steroid
products and stability of the system.
Optimizing some conditions for spray drying in synbiotic capsule from Bacillu...inventionjournals
The study is of determination of various conditions for spray drying in producing synbiotic in the form of capsule from Bacillus subtilis natto. The experiments were conducted to examine effects of various factors such as the resistant starch-to-matodextrin ratio before drying, the inlet gas temperature and the inlet flow. The optimization experiments are administered: a ratio of wall materials to the core material is 5% (w/v) in which the ratio of resistant starch and maltodextrin is 1:9, the inlet gas temperatureis 1100C, the inlet flow is 5.60 ml/minute and the spray pressure is 2 bar. In the condition for spray drying as mentioned earlier, the initial step for trial production of synbiotic capsules was conducted with the cell density of B.subtilis natto being 8.55 ± 0.18 log(CFU/g), the activating effect of nattokinase is 518.2 FU/g and the moisture is 9.11%. During 60 days’ preservation of the product, the resulting indexes such as the cell density of B.subtilis natto in the capsule, the activating effect of nattokinase and the moisture are stable.
Validation Of Radiation Sterilization Dose For Proteases Immobilized On Aldeh...iosrphr_editor
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
Comparison of Three Slaughtering Methods of Goat on Carcass and Prime Cuts Re...inventionjournals
Procedures employed on slaughtering of goat varies due to lack of information on the effect of each process such as scalding, flaying and singeing on the different slaughter parameters and prime cut recovery. Using fifteen male Anglo-nubian goats at 8months old, the effect on slaughtering weight, hot carcass, chilled carcass, dressing percentage and drip loss as well as in the prime cut production was documented. Results show that on slaughter weight, there is no significant difference obtained due to the uniform management production system used in raising the animals. On hot and chilled carcass weights, significant difference was found (p=0.01). Difference on dressing percentage revealed that singeing has the highest recovery rate at 59.92%, and values obtained is significantly different (p=0.05). Drip loss is also highly significant (p=0.05). On the prime cut recovery, highly significant differences were obtained on percentage (%) shoulder, loin and flank, offals and the skin and other cuts did not vary significantly from each other. It can therefore be concluded that each of procedures employed has an effectto the differentparameters considered in goat slaughtering and in prime cut recovery.
Cell-free DNA Levels Serum Patients with Benign and Malignant Epithelial Ovar...inventionjournals
An elevated level of cell-free DNA (cfDNA) in the blood circulation has detected in cancer patients in comparison with healthy controls. CfDNA circulation in plasma and serum extensively studied and the results are highly variable due to many factors influence the test results that was preanalytic factors as well as analytic factors. Objectives: Is there any difference in the concentration of serum DNA among patients with benign epithelial ovarian tumors and malignant epithelial ovarian tumors? What is the clinicopathological variable that influences the cfDNA circulation? Method: Venous blood drawn with plain vacutainer, centrifuged at 1,000 rpm for 30 minutes, serum kept in -800 C freezer. The cfDNA extracted used NaI method.Results: Collected 30 cases of the benign ovarian tumor and 54 cases of malignant ovarian tumors. The average level serum cfDNA of benign epithelial ovarian tumors and malignant epithelial ovarian tumors were 24.6 ng/mL and 22:29 ng/mL respectively and statistically was not significantly different (p = 0.64). In multivariable analysis with linear regression, there were no clinicopathological variables that statistically significant influence the cfDNA levels in patients with epithelial ovarian tumors where p > 0.05. Conclusion: Concentration of cfDNA circulation of benign epithelial ovarian tumors a little bit higher than malignant epithelial ovarian tumors, but statistically was not significantly different. There was no clinicopathological variable influence the concentration of cfDNA circulation of ovarian tumors.
No, Orally Administered Hemolymph Of Limicolaria Aurora Does Not Reduce Blood...inventionjournals
The oral administration of the hemolymph of Limicolaria aurora at doses of 22.8 and 45.6 mg/kg body wt did not result in any significant difference in the systolic, diastolic, pulse pressure, mean arterial pressure and heart rate of both normotensive and adrenaline induced hypertensive wistar rats. The data from this study show the lack of antihypertensive potential associated with the oral ingestion of Limicolaria aurora hemolymph.
Assessment of Microbial Contamination of the Tooth Brush Head Used On Orthodo...inventionjournals
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Dual Spaces of Generalized Cesaro Sequence Space and Related Matrix Mappinginventionjournals
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Isolation, Screening and Selection of Fungal Strains for Potential Cellulase ...inventionjournals
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Some Properties of M-projective Curvature Tensor on Generalized Sasakian-Spac...inventionjournals
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Co- Movements of India’s Stock Market with Bond Market and Select Global Stoc...inventionjournals
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IJPCBS 2012, 2(1), 110-116 Kavya et al. ISSN: 2249-9504
110
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Available online at www.ijpcbs.com
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Optimization of Bacillus Subtilis Natto Immobilization Process on Alginate – Chitosan Complex and Its Application for Nattokinase Fermentation
1. International Journal of Pharmaceutical Science Invention
ISSN (Online): 2319 – 6718, ISSN (Print): 2319 – 670X
www.ijpsi.org Volume 5 Issue 3 ‖ June 2016 ‖ PP.25-30
www.ijpsi.org 25 | P a g e
Optimization of Bacillus Subtilis Natto Immobilization Process on
Alginate – Chitosan Complex and Its Application for Nattokinase
Fermentation
Hien T. Ho1
, Huong T. Nguyen2
1,2
(Department Of Biotechnology – Ho Chi Minh City University Of Technology)
ABSTRACT: Nattokinase is a potent fibrinolytic enzyme with the potential for fighting cardiovascular diseases.
In this study, Bacillus subtilis natto were immobilized in the alginate – chitosan complex for fermentation of
nattokinase enzyme. Six factors affecting the efficiency of immobilization cells were screened by Plackett –
Burman design including: concentration of alginate, concentration of chitosan, pH of chitosan, concentration of
CaCl2, added cells density, shaking time after supplementing chitosan. Results of optimization have identified
two factors affecting the efficiency of cell immobilization. They are concentration of alginate (2.5%) and added
cells density (approximately 5.86 million colonies per milliliter). With these two factors optimized and others
kept at the normal level, immobilization efficiency reached 90.73%. After Bacillus subtilis natto had been
immobilized by optimization of parameters, we conducted application for fermenting nattokinase. For 24 hours
of fermentation, nattokinase enzyme activity reached 71.80 ± 0.19 FU/ml. Immobilized Bacillus subtilis natto
cells were reused 6 times and on the 6th
time of reuse, nattokinase enzyme activity only decreased 2.7% in
compared with the 1st
reuse.
Keywords: Bacillus subtilis natto, immobilized cells, alginate, chitosan, nattokinase
I. Introduction
Nattokinase [EC 3.4.21] is an extracellular serine protease, this enzyme was demonstrated being a fibrinolytic
enzyme and considered a potential agent in the effective treatment of obstructive vascular disease [1]. This
enzyme has been discovered from various food sources such as Natto of Japanese [1], Doen-jang of Korea or
Dauchi of Chinese [2] and from various microbial especially the strains Bacillus subtilis bacteria in traditional
fermented foods from soybean [3]. Bacillus subtilis natto strains are capable of the highest yield of nattokinase
biosynthesis.
The technique to immobilize cells and enzyme by entrapment in matrix method has been applied for high
efficiency in the food industrial, pharmaceutical and environmental treatment [4]. Among the carriers applied in
the immobilization process, alginate and chitosan are widely used because these carriers are natural polymers
and have some advantages: Entrapment within gels of natural polymer is very mild technique and damage to
living cells can be minimized, easily handling, inexpensive and safe for users [5]. However, the immobilization
of cells in alginate gel has the disadvantage that immobilized bacteria are surrounded by a gel network which
strongly limits their movement. When growth occurs, bacteria push the gel network away and colonies
containing densely packed bacteria are formed. As the colony expands, it may eventually reach the surface of
the gel bead. This may lead to an eruption of the colony to the surrounding medium and limiting the reuse.
Moreover, alginate gel beads are unstable in the medium containing citrate and phosphate that are often used as
a buffer solution in the fermentation broth. Besides, the immobilized of cells in chitosan has some disadvantage
due to the small gel pore size which limits the diffusion of nutrients and metabolic products between the
fermentation broth and cells in the gel bead. In order to overcome this limitation, bacteria will be entrapped in
calcium alginate gel beads, after that they are coated with chitosan. The combination of the amino group of
chitosan and carbonate group of alginate will make up the gel network strength, which will help reduce cells
released and increase the number of cell reuse [6,7]. In addition to that, chitosan’s characteristics of antibacterial
in the fermentation broth and stability at high temperatures will create favorable conditions for Bacillus subtilis
natto cells to grow strongly [8].
II. Material And Method
1.1 Microorganism Strain And Cultural Medium
Bacillus subtilis natto was selected from bacteria collection of Ho Chi Minh University of Technology.
Bacillus subtilis natto was cultured in the nutrition broth (NB) and nutrition agar (NA), the pH 7.5, the
temperature at 370
C, the agitation of 200 rpm.
Medium for fermentation broth composed of Glucose 5.6 (g/l), Peptone 13 (g/l), MgSO4.7H2O 0.875 (g/l), NaCl
2.5 (g/l), CaCl2 0.025 (g/l) [9].
2. Optimization Of Bacillus Subtilis Natto Immobilization Process On Alginate – Chitosan Complex..
www.ijpsi.org 26 | P a g e
Six factors effecting on efficiency of immobilization: concentration of alginate (1.5-3.5%), concentration of
chitosan (0.2-0.6%), pH of chitosan (5-6); concentration of CaCl2 (1-3%), initial supplement cells density (10 -
100 million colony forming units per milliliter); shaking time when supplementing chitosan (20-30 minutes)
1.2 Methods
2.2.1 Optimization process of Bacillus subtilis natto
- Screening the factors effecting on the efficiency immobilization of cell.
Six variables were examined in the Plackett Burman matrix with different 12 runs. We determined the
performance of immobilization for each validation formula and analyzed the factors having great impacts on
efficiency of immobilization cells. The main effecting factors had p value < 0.05. The chosen factors were
designed to determine the relationship between those and the response.
- Optimizing parameter for cells immobilization process
With the selected factors from the screening experiments, we carried out the initial experiment with original
values (-1,+1). Based on “Lack of fit” test, we determine the relationship between the chosen factors and
response. After analyzing the initial experiments, we determined whether the factors having great impacts on the
high regression equation are suitable or not. If the regression is linear function, we design the steepest
experiments to identify the suitable range of factors.
Based on that, we conducted the experiments for response surface methodology having the central composite
designs and determined function of the poly - nominal regression accurately to describe relationship between the
efficiency of immobilization and each factors.
The optimized results of response surface methodology were calculated by software simulation to determine the
highest immobilization efficiency and immobilization conditions.
Analyzing data and identifying the regression were carried out by Minitab 16.
2.2.2 Using immobilization of cells for fermentation nattokinase
We performed immobilization of cells with optimal parameters found from optimizing experiment then
conducted fermenting enzyme nattokinase.
Immobilized cells were fermentation with: broth being medium for fermentation, temperature at 370
C, agitation
at 200 rpm. The survey conducted on process of enzyme nattokinase biosynthesis was based on time of
fermentation by immobilization of cells. Survey process started at 8th
hour of fermentation and on every 4 hours,
sample was taken to determine enzyme nattokinase activity. Fermentation was stopped at 28th
hour.
Survey reused immobilized cells by stopping fermentation at 24th
hour, collecting broth for determine
nattokinase activity and transferring immobilized cells to fresh broth medium.
2.2.3 Analyzing nattokinase activity
Nattokinase activity was determined by ability to hydrolyze fibrin fiber. Bacillus subtilis natto fermentation was
stopped after 20 hours and broth was centrifuged at 13000 rpm for 20 minutes and obtained the supernatant to
determine nattokinase activity.
Tris-HCl (50mM, pH 7.5) of 1.3 mL and 0.4 mL 0f 0.72% (w/v) fibrinogen solution were taken in vials and kept
in water bath (370
C) for 5 minutes. Then 0.1 mL thrombin (20U/mL) was added and kept in water bath (370
C)
for 10 minutes. To this clot, 0.1 mL of enzyme was added. After incubation (370
C, 60 minutes), 2 mL of 0.2 M
trichloroacetic acid (TCA) was added. Vials were kept 20 minutes and centrifuged at 3000 x g for 5 minutes.
One unit enzyme activity is defined as the amount of enzyme required to produce an increase in absorbance
equal to 0.01 in 60 minutes at 280 nm [10].
III. Result And Discussion
1.3 Screening main effective factors of the cell immobilization efficiency
The cell immobilization efficiency and the effective values obtained from Plackett Burman matrix by
experiments were illustrated in Table1. Two factors notably influenced in cell immobilization efficiency
(p<0.05) were alginate concentration and cell density.
The alginate concentration factor and the added cells density had effect value (39.9, p = 0.01) and (45.56, 0.01)
respectively. The remaining factors had not effect significantly in designed experiments.
Table 1: The factor in Plackett Burman matrix and its effect to immobilization cells
Name of factors Symbols
of factor
Values of factors Main
effect
P values
Low (-1) High (+1)
Alginate concentration (%) X1 1.5 3.5 39.9 0.01
Chitosan concentration (%) X2 0.2 0.6 0.1 0.767
pH chitosan X3 5 6 0.09 0.774
CaCl2 concentration (%) X4 1 3 5.61 0.064
The added cells density (million X5 10 100 45.56 0.01
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colonies/ml)
Shaking time X6 20 40 5.75 0.062
R-sq = 95.10%
1.4 The optimal values of parameters in immobilizing cell process for highest efficiency of immobilization
We determined the relationship between cell immobilization efficiency and two main effective factors through 9
initial experiments. Through initial experiments, we found that alginate concentration has greatest impact on
immobilization efficiency than added cell. Through the levels of two variable and their effect on immobilization
efficiency (Table 2) we have some following remarks: Specifically, alginate concentration had main effect 165.2
and P value 0.00 then added cell had main effect 31.65 and P value 0.002. The analyzing data demonstrated R-
sq = 98.63% and Lack of fit test having p-value of 0.05, this meant that the immobilization efficiency and two
main effective factors were not in the linear relationship.
Table 2: The affecting level of two factors
Name of
factor
Symbol Value of factor Main effect P
valueLow (-1) Central (0) High (+1)
Alginate
concentration
(%)
X1 1.5 2.5 3.5 165.02 0.000
Cell added
(million
colonies/ml
X5 10 55 100 31.65 0.002
We identified the function of the poly-nominal regression accurately. The experiment for response surface
methodology having the central composite designs (RMS-CCD) with the main effective factors (-α, -1, 0. +1
+α) were conducted through 13 experiments, including 5 central experiments, 4 corner experiments and 4 axial
experiments.
The highest cell immobilization efficiency was the 4th
run (X1=0, X5= -α) of all experiments, which reached
90.73%. The lowest cell immobilization efficiency was the 3rd
run (X1= -1, X5= 1) which reached 30.23%, the
efficiency of immobilization. The experiments were illustrated in Table 3.
Table 3: The efficiency of immobilization Bacillus subtilis natto by alginate – chitosan complex in central
composite designs.
Run Code value Un code value Efficiency
Y (%)X1 X5 X1
(%)
X5
(million colony/ ml)
1 0 0 2.5 55 75.67
2 -1 -1 1,5 10 39.76
3 -1 1 1.5 100 30.23
4 0 -1.41 2.5 5.858 90.73
5 0 0 2,5 55 74,01
6 -1.41 0 1.085 55 32,41
7 1.41 0 3.914 55 45.56
8 0 0 2.5 55 76.58
9 1 1 3.5 100 59.32
10 1 -1 3.5 10 80.27
11 0 0 2.5 55 75.85
12 0 +1.41 2.5 104.142 42.71
13 0 0 2.5 55 76.36
R-sq – 90.15%
In this case, we determined the relationship between the main effective factors and response. The poly-nominal
regression equation was determined below.
Y (%) = -67.86 + 106.86*X1 + 1.35*X5 – 18.47*X1
2
- 2.27*X5
2
– 6.34*X1*X5
In above equation, Y (%) is symbol for cell immobilization efficiency, X1 (%) is symbol for alginate
concentration, X5 (million colonies per milliliter) is symbol for added cell density.
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Figure 1: The equation contours show the relationship between Y (%) and X1, X5
When added cells density was above central value, immobilization efficiency was reduced. This shows that,
added cells density being higher than central value will lead to competitive nutrients of cells in the gel bead.
Besides, the increasing by-product in the fermentation medium leads to inhibition of cell grows. Therefore, cells
enter death phase and the cells density in the gel bead decreases.
When alginate concentration increased above the central value, the immobilization efficiency also increased.
However, when alginate concentration reached value +1, despite the increasing in alginate of concentration the
immobilization efficiency was not increased.
The reason is that when alginate concentration increases, the viscosity increases too, so the ability of diffusing
cells into alginate suspensions is limited. On the other hand, when alginate concentration decreases below the
central value, the immobilization efficiency also decreases because low alginate concentration lead to the large
size pore gel, cells in gel bead are easily released to fermentation medium.
The maximum efficiency of immobilization Bacillus subtilis natto on alginate – chitosan complex was 90.73%
when alginate concentration is 2.5% and added cell density is 8.58 million colonies per milliliter, the others
factors remaining at central level.
1.5 Using immobilization Bacillus subtilis natto for fermentation of nattokinase
1.5.1 Investigating biosynthesis process of nattokinase enzyme based on time of fermentation.
Survey time period started at 8th
hour and stopped at 28th
hour of fermentation. The nattokinase enzyme activity
obtained increased as survey time increased. At the time of 8th
hour after fermentation, nattokinase enzyme
activity reached 31.46 FU/ml because in this stage, cells was in log phase and using nutrition for growing. The
results of survey process were illustrated in the figure 2.
Figure 2: The effect of fermenting time on biosynthesis of nattokinase enzyme process.
After 20 hours of fermentation, nattokinase enzyme activity reached 67.21 FU/ml and increased 2.2 fold
compared with at the 8th
hour of fermentation. The time from 24th
hour to 28th
hour correspond with the time
when cells in the stationary phase so nattokinase enzyme activity just slightly increased by 0.16 FU/ml. This
indicated that the increase in density of cells in the fermenting medium is proportional with obtained nattokinase
enzyme activity. At 28th
hour of fermentation nattokinase enzyme activity reached maximum 71.96 FU/ml.
However for the 4 hours prolonged time, nattokinase enzyme activity only increased 0.16 FU/ml having no
economic effect. Because the purpose of fermenting nattokinase enzyme by immobilized cells is to obtain the
highest enzyme activity and cell density, we chose the time to stop fermentation at the 24th
hour.
1.5.2 Comparing nattokinase enzyme activity obtained from fermenting process using free cells and
fermenting process using immobilized cells.
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From the investigating results obtained from biosynthesis of nattokinase enzyme process based on fermentation
time, we conducted fermenting immobilized cells and fermenting free cells with the same conditions. The result
was illustrated in figure 3.
Figure 3: Comparing nattokinase enzyme activity value obtaining from fermenting free cells and fermenting
immobilized cells.
For the free cells experiment, the maximum nattokinase enzyme activity obtained at 20th
hour of fermentation
was 69.13 FU/ml. Meanwhile, the maximum nattokinase enzyme activity obtained from fermenting the
immobilized cells was 71.96 FU/ml at 28th
hour of fermentation. The cause of the difference is that immobilized
cells take longer time to adapt with fermentation medium than free cells do. With free cell, when cells were
added in to fermentation broth, they immediately and directly contacted with the substrate. As for the
immobilized cells, cells were locked up behind the barrier layer of particle gel so cells used substrate after being
diffused through the membrane. On the other hand, the diffusion also has the limitation because it is effected by
concentration of substrate and size of the substrate. However for free cell, when increase time for fermented
from 20th
hour to 28th
hour, the nattokinase enzyme activity obtained was not increase. While the nattokinase
enzyme activity obtained by the immobilized cell was increased 4.75 FU/ml.
1.5.3 Investigating nattokinase enzyme activity in a series of immobilized cells reuse
We conducted survey nattokinase enzyme activity for a number of immobilized cells reuse times. In every 24
hours, we stopped the fermentation, transferred gel beads to fresh broth and continued fermenting process with
the same condition on the fresh broth. The results of investigating nattokinase enzyme activity over time of
reuse were illustrated in the figure 4.
Figure 4: Survey reusing the immobilized cells
The nattokinase enzyme activity in the first reuse reached 73.13 FU/ml, the highest level among the reuse times
and higher than the initial of immobilized cells. Particularly, the nattokinase enzyme activity obtained in the first
reuse and the second reuse were higher than the initial of immobilized cells by 1.33 FU/ml and 0.61 FU/ml
respectively. The cause of this difference is due to when we stopped process of fermented at the time of 24th
hour, the cells in gel beads are ungrowing hormogenous. Some are growing, others might be in the process of
biosynthesis of nattokinase enzyme thus nattokinase enzyme activity obtained was not in the highest level at the
initial fermentation of the immobilized cells. At the time of the first reuse nattokinase enzyme activity obtained
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reached the highest level because cells in the gel bead are not undergo growth stages which mainly using
nutrition for the biosynthesis of nattokinase enzyme process. At the next time reuses, obtained nattokinase
enzyme activity decreasing compared with the previous reuses may be due to after each reuse, a number of cells
were being released in to fermentation broth. So cells density in the gel beads were decreased and obtained
nattokinase enzyme activity were also decreased.
However after reusing for 6 times, obtained nattokinase enzyme activity compared with the 1st
time of reuse is
decrease by 2.73 FU/ml, this reduction is not significant. The survey result suggests that alginate – chitosan
complex had its effect in reducing cells release and increasing times of reuse.
IV. Conclusion
By the response surface methodology, we had investigated two factors affecting the immobilization efficiency
are: alginate concentration and added cell density. The Optimized of parameters for six factors are concentration
of alginate (2.5%), concentration of chitosan (0.4%), pH of chitosan (5.5), concentration of CaCl2 (2%), added
cell density (approximately 8.58 colonies per milliliter) and shaking time when supplement chitosan (30
minutes). The highest of immobilization efficiency reached 90.73%.
The immobilized cells were reused 6 times and obtained nattokinase enzyme activity in the 1st
time of reuse and
in the 6th
of reuse are 73.13 ± 0.14 FU/ml and 70.40 ± 0.09 FU/ml respectively.
Acknowledgment
We would like to full appreciate the help of the professors/teachers in the Chemistry Faculty of Ho Chi Minh
University of technology in this study. This review is funded by Vietnam National University Ho Ch iMinh City
(VNU-HCM) under grant number C2016-20-31.
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