One-Day National Conference on Innovations in Biological Sciences On 10 January 2020 in the Department of Biosciences, Saurashtra University, Rajkot, Gujarat, India Abstracts of the papers presented in this conference by the members of Prof. Satya P. Singh Lab

1. One-Day National
Conference on Innovations
in Biological Sciences
On 10 January 2020 in the Department of
Biosciences, Saurashtra University, Rajkot,
Gujarat, India

6. Prof. Satya P. Singh, PhD
UGC-BSR Faculty Fellow (Professor-Emeritus)
UGC-CAS Department of Biosciences
Saurashtra University
RAJKOT-360 005. Gujarat, INDIA
Head, Department of Biosciences, Saurashtra University (2003-2020)
Coordinator, UGC-CAS Program, Saurashtra University (2013-2018)
Coordinator, DST-FIST Program (2007-2012)
Coordinator/PI, DBT-Multi Institutional Project (2007-2012)
PI, MoES-Multi Institutional Net Working Project (2014-2018)
Coordinator, M.Sc. Biotechnology, Saurashtra University (2004-2017)
Coordinator, UGC DSA-Phase-3 and UGC COCIST (2003-2006)
Email: satyapsingh@yahoo.co satyapsingh125@gmail.com
spsingh@sauuni.ac.in
SU Website URL: saurashtrauniversity.edu/university/academic-
departments/department-of-biosciences/staff/dr-satya-prakash-singh/

7. ResearchGate: https://www.researchgate.net/profile/Satya_Singh5
GoogleScholar: https://scholar.google.com/citations?hl=en&user=jiAzOcg
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UGC: https://vidwan.inflibnet.ac.in//profile/68903/Njg5MDM%3D
Abstracts of the papers presented from
Prof. Satya P. Singh’s Laboratory
Category: PhD
Gene profiling, phylogeny and structural features of alkaline proteases of
Haloalkaliphilic bacteria from Little Rann of Kutch
Hitarth B. Bhatt and Satya P. Singh*
UGC-CAS Department of Biosciences, Saurashtra University, Gujarat, India -360005
Email: satyapsingh@yahoo.com; hitarth_bhatt@yahoo.co.in
Abstract
The Little Rann of Kutch is a saline desert with unique ecosystem and yet unexplored for
microbiological studies. In addition, this is the first report on the gene profiling, phylogeny and
structural properties of alkaline proteases of the haloalkaliphilic bacteria of this habitat.
Alkaline proteases genes of 19 different protease producing bacteria were amplified using 12
different primer pairs. Among the protease producing haloalkaliphilic bacteria, maximum
number of amplifications were achieved with primer BPAP2 (8 strains) followed by SPSAP
and SPSBH (7 strains), BPAP1, BLAP, BAAP and SPSVB (4 strains), PCAP and GMAP (1
strain) primers. However, 3 primers: BLIAP, BLIAP2 and SPS-6 failed to amplify protease
genes in any of the examined strains. The amplification profile indicated the diversity of the
protease genes with respect to the size of the amplified genes. The amplified products ranged
from ~ 400 bp to ~ 1700 bp. BLASTn analysis revealed that all protease coding genes belonged
to the peptidase S8 family. Further, sequence similarity ranged between 84.49- 99.53%. A total
of 8 amplified products were sequenced and subjected to phylogenetic analysis. Phylogenetic
analysis revealed cladding pattern of alkaline protease genes in four separate clusters indicating
considerable diversity, while the structural analysis established the thermostability and

8. halostability of the alkaline proteases. The genes of high predicted stability would be cloned in
mesophilic host to extend the study further.
Key words: Alkaline protease; Gene profiling, Little Rann of Kutch; Haloalkaliphilic bacteria;
Gradient PCR; Structure and function relationship; Phylogeny; Structure to function
relationship
Some aspects of it now published as below:
Bhatt, H.B. and Singh S.P. 2020, Cloning, Expression and structural elucidation of a
biotechnologically potential alkaline serine protease from a newly isolated Haloalkaliphilic Bacillus
lehensis JO-26, Frontiers in Microbiology (IF 4.25), 11:1-16,
https://doi.org/10.3389/fmicb.2020.00941
Isolation and characterization of Plant Growth Promoting Rhizospheric bacteria from
Limonium stocksii
Paragi Jadav, Kalpna Rakholiya, Mital Kaneria, S.P. Singh
Department of Biosciences, Saurashtra University, Rajkot-360005, Gujarat, India
E-mail: paragijadav@gmail.com
Abstract
Plant Growth Promoting Rhizospheric bacteria (PGPR) are beneficial microbes that can
enhance plant growth against biotic and abiotic factors. The present investigation was carried
out to isolate and characterize the PGPR from the rhizospheric region of Limonium stocksii, a
halophyte. Salt tolerance test for the organisms was performed up to 15% of salt
concentration. Out of 10 bacterial isolates, 06 were able to grow at 15% salt concentration.
The isolates were further investigated for their PGP traits and secretion of the hydrolytic
enzymes. Among them, six isolates showing noticeable efficacy were selected for further
studies. The ongoing work would establish the usefulness of these organisms to enhance plant
growth via diverse mechanisms eventually leading to the reduction and/or replacement of the
synthetic fertilizers and pesticides in agriculture practices
Keywords: Limonium stocksii, Halophyte, PGPR, Enzymes, Salt tolerance

9. Sequence based analysis of the non-cultivable actinomycetes of the saline habitats of
coastal Gujarat near Dwarka
*Kruti G. Dangar and Satya P. Singh
Department of Biosciences, Saurashtra University, Rajkot, Gujarat, India
*Currently associated with- Department of Biochemistry, Saurashtra University, Rajkot,
Gujarat, India
Email: satyapsingh@yahoo.com
The diversity and identification of the non-cultivable actinomycetes of the saline
habitats have been investigated using metagenomic approach. The sequence based
metagenomic analysis provided detailed information about the diversity, phylogeny
and putative physiological functions of the actinomycetes in the saline habitats. The
diversity of the organisms was analyzed by amplification and sequencing of the 16Sr
RNA genes of the total genomic DNA isolated form the salt enriched soil. The
metabolic functions of the organisms were also analyzed. Microbial classification
based on the OTUs represented Bacteria belonging to various phylum. The relative
abundance of the actinomycetes in the studied habitats was assessed at the level of 0.1%.
The dominant domain consisted of the phylum Bacteroidetes (37.9%) and Proteobacteria
(22.0%) and Gemmatimona (36.4%). The results showed that composition of the
microorganisms of the saline ecosystem reflects the function in the saline habitat. The
level of the microbial diversity was substantially greater than that of the laboratory
cultivable isolates.
Key Words: Metagenomics, saline habitats, non-cultivable acinomycetes,

10. Category: M.Sc.
Statistical Optimization of the production of Amylase and
Protease in marine actinomycetes using Plackett-Berman
design
Vipul Lagariya, Dalip S. Rathore and Satya P. Singh⃰
UGC-CAS Department of Biosciences, Saurashtra University, Rajkot, Gujarat, India- 360005
Email: lagariya01@gmail.com satyapsingh@yahoo.com
Abstract:
In this study, amylase and protease were co-produced by a marine actinomycete,
Nocardiopsis alba KaM14. Initially the parameters for the production of these
enzymes were screened by one variable at a time approach (OVAT). In order to
get further insight into the enzyme production, the fractional factorial design,
namely Plackett-Berman design was implemented. Seven parameters; Starch,
Gelatin, Peptone, Yeast extract, pH, NaCl concentration and Temperature were
taken into consideration for the optimization. The amylase was assayed using
DNSA method and protease assay was based on the Anson -Hagihara method.
The starch and NaCl concentrations significantly influenced the amylase
production, while yeast extract affected the production of protease. The study
signifies the reduction in the cost of co-enzyme production.
Keywords: Co-production of enzymes, Halophilic actinomycetes, Plackett-
Berman Design

11. Category: M.Phil.
Production and Purification of amylase from the marine
actinomycetes Nocardiopsis alba
Ankita Dobariya and Satya P. Singh⃰
Email: ankitadobariya108@gmail.com ; satyapsingh@yahoo.com
UGC-CAS Department of Biosciences, Saurashtra University, Rajkot, Gujarat, India
Halophilic amylases are in demand due to their suitability in extreme environmental conditions.
The present study is based on the production and purification of an amylase a from marine
actinomycete Nocardiopsis alba. Different parameters, such as pH, NaCl concentrations,
substrate concentrations and incubation time were evaluated and optimized using ‘One variable
at a time method (OVAT)’. The amylase was secreted in significantly high quantity and the
enzyme activity was measured by DNSA method. Further, the partial purification of amylase
was achieved using chilled acetone precipitation. Ammonium sulphate, Methanol and PEG
(Poly Ethylene Glycol) mediated partial purification was not successful. The enzyme was
further purified by Size Exclusion Chromatography (SEC) rather than Hydrophobic Interaction
Chromatography (HIC). The purified enzyme was analyzed on SDS PAGE for the assessment
of the purity and determination of the molecular mass.
Keywords: Marine actinomycetes, Amylase, OVAT, Acetone Precipitation and Size
Exclusion Chromatography (SEC)
Category: PhD
STATISTICAL OPTIMIZATION OF THE CO-PRODUCTION OF
AMYLASE AND PROTEASE FROM HALOALKALIPHILIC
ACTINOMYCETES, STREPTOMYCES LOPNURENSIS
Dalip Singh Rathore and Satya P. Singh
Email: rdalipsingh21@gmail.com ; satyapsingh@yahoo.com
UGC-CAS Department of Biosciences, Saurashtra University, Rajkot, Gujarat, India
Actinomycetes are potential source of industrially viable products such as bioactive compounds
and enzymes. Amylase and protease are potentially useful in a wide range of applications in
various fields. Individual production of amylase and protease adds to the cost of fermentation.
Therefore, the present study has focused on the co-production of amylase and protease from a
marine actinomycete, Streptomyces lopnurensis. Initially, the parameters affecting the co-
production were adjudged based on ‘One Variable at a Time’ (OVAT) approach. In order to
gain further insight into the production and optimization, statistical methods; Placket Berman

12. Design (PBD) and Box-Behnken Design (BBD) using Minitab 14.0 were used. The data was
further analyzed to examine the statistical fitness and validity by regression equation. The
predicted values were statistically significant with the observed values. A mutual repression of
amylase and protease was observed in co-production. The enzyme secretion was initially
growth dependent. However, it gradually turned into growth-independent. Initially, amylase
was produced in higher quantity as compared to protease. However, after three days of growth,
protease production increased suppressing the amylase production. The media used for the co-
production was economically viable as it contained easily available and cheap nutrients. The
optimization of co-production of two enzymes by statistical methods assumes significance in
the larger context of using enzymes from extremophiles.
Keywords: Streptomyces lopnurensis, OVAT, Placket-Berman Design(PBD), Box-Behnken
Design (BBD) , Co-production and Growth dependent enzyme secretion
Now Published as below:
Rathore, D R and Singh, S.P. 2021. Kinetics of growth and co-production of amylase and protease in
novel marine actinomycete, Streptomyces lopnurensis KaM5. Folia Microbiologica (Springer; IF:
1.70), https://doi.org/10.1007/s12223-020-00843-z
Category: Research Scholar
Abstract
Screening and production of Extracellular proteases of marine actinomycetes from
Alang, Gujarat coast
Mahejbin A. Sheikh, Dalip Singh. I. Rathore, Satya P. Singh
Emails: mahejbin.sheikh92@gmail.com, satyapsingh@yahoo.com
UGC-CAS Department of Biosciences, Saurashtra University Rajkot, Gujarat, India
Marine actinobacteria are ecologically and biotechnologically valuable microorganisms.
Beside antibiotics, they have potential to produce many commercially important bioactive
compounds and industrially useful enzymes. Sixty six seasonal marine actinomycetes
discussed in this study were screened and found to secrete extracellular proteases and amylases.
Among them, 14 isolates were selected for the protease enzyme production in liquid culture
media. Effect of salt concentrations on growth and protease production were studied for the
selected three potent isolates for 16 days. The results indicated that the production of protease
in all isolates was optimum during stationary phase of growth with 5% salt concentration. The
ability of the actinomycetes to grow up to 20% salt concentration indicated their halophilic

13. nature. Overall, the study reflects the biocatalytic potential of the marine actinomycetes from
Alang, Bhavnagar Coast, Gujarat.
Keyword: Marine Actinomycetes, Halophiles , Biocatalytic potential, Extracellular Protease
Construction and analysis of marine sediment metagenomic library to mine biocatalysts
Nirali M. Raiyani and Satya P. Singh
niraliraiyani6@gmail.com, satyapsingh@yaoo.com
UGC-CAS Department of Biosciences, Saurashtra University, Rajkot 360 005, Gujarat, India
Metagenomics is a powerful tool that allows culture-independent analysis of complex
microbial communities of a given habitat. Since 95-99% of the microorganisms in majority of
the habitats are uncultivable, it significantly limits the information on the genomes and genes
encoded. Functional screening of the metagenomic DNA libraries is a promising approach to
search for genes encoding novel enzymes. As per the current estimation, the soil microbiome
is considered as one of the most diverse and complex microbial ecosystems. In the present
study, direct extraction methods were developed for the extraction of metagenomic DNA based
on physical, chemical and enzymatic methods in various combinations. The metagenomic
DNA extracted from the marine sediment was used for construction of metagenomic library.
40 kb size metagenomic DNA was ligated with pCC1FOS fosmid vector and packaged into
lambda bacteriophage followed by the transfection of EPI300T1R
host strain. The large insert
metagenomic library was constructed and the analysis of the clones for different biocatalysts;
such as amylase, protease and lipase is currently being investigated.
Key words: Uncultivable microorganisms, Marine sediment, Metagenomic DNA,
Metagenomic library, Functional screening
Related Aspects Published as below:
Raiyani, Nirali and Singh S.P. 2020, Extraction of environmental DNA, construction of metagenomic
libraries and functional screening of enzymes from salt pan soil, Indian Journal of Geo-Marine
Sciences, Accepted (NISCARE, CSIR, IF 0.50),
Raiyani, Nirali and Singh S.P. 2020, Taxonomic and functional profiling of the microbial communities
of Arabian Sea: A Metagenomics approach
Journal: Genomics (Elsevier, IF 6.20), 112:4361- 4369 https://doi.org/10.1016/j.ygeno.2020.07.024