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1



1




      ––––




    1870
2
3




(fractional distillation)
       2
4




1.




     (Operational Definition)
      -


                                6
5




-
-
-



    -




        -
6




2
7




                 (   )




1.
     2   -   3
8




2.



3.
4.



5.



6.
                                    2

                     2-3


                               6



     1.5   –   5.0

                                   2-5

                           1
9



2.1.1




        1.




        2.




             45%
10




            400

        6

        6

        6

            6

            6

        2

2.2

2.2.1

        (   )




        Amylomyces   sp.
11




Aspergillus   sp.                       Rhizopus          sp.


Mucor                Saccharomycopsis sp.          Pichia sp.

                    Saccharomyces sp.




                                                   +



                                    +




2.2.2
12




                C6H12O6 - - - > 2 C2H5OH + 2CO2




                C6H12O6 + 6O2 - - - > 6CO2 + H2O +



2.3




      2.3.1                                        simple
distillation)
      2.3.2            (fractional distillation)
      2.3.3
13



      2.3.1                                 simple
distillation)

                                simple distillation)



                   80




              1.
              2.        boiling stone       boiling
      chips
              3.
              4.
14



    5.


              2-3


    6.




    1.


    2.
    3.
    4.




    2.3.2   (fractional distillation)


2
15
16




2.3.3




2.4


        2




            100
17



                   78.3


                   10%         93


                              55%


              82               %




78




     78   –        85
                               90


                     (    )
18



2.4.1


  1.


        simple   distillation)



                       (    )
19


  2.



(fractional   distillation)
20




                                 (2                          )
                                                            %



                            Saccharomyces cerevisae
 x           /



                                                  modified
Lane-Eynon                                                  %
                                                          HPLC




                 %      ,                 %                 %
                                                            %
                        %             %


                                                             -


Enterobacteraerogenes
                                                      -
                                                  (CGWO)
                                                      37    oC
21



                  Conversion
(                                                          )
                                       (            30oC)



                                                           Conversion


CGWO                               Conversion                            -
                                                                         -
             30 g/l




                                                       4
              1                                 (      .                 )
             2,    3           4
Saccharomyces cerevisiae NP 01, S. bayanus EC 1118                      S.
cerevisiae        Sweden
                                                                        2
                                                35                 32
                                           10




                                   2                           -
                                                      500
22



                                                 5
                                          Z.                 mobilis




5


                     Z.      mobilis           TISTR            548



                100                                      -     (pH)
               5.5                              100
                                                              0.463



                          NH4Cl                      1

                                                              0.419



    Altintas


Saccharomyces                 cerevisae                      YPB-G


                                                 4
23



                                          0.118              0.183



      Dung

                                                                  29


                 119                                   53

                 15                               51

                             Amylomy          ces             rouxii

                            Saccharomyces cerevisiae

      Tsuyoshi

                                                  6

                                         20

                                          4                  1

Saccharomyces bayanus          2     Candida glabrata         3

Pichia anomala          4          Saccharomycopsis fibuligera




1,2      3

      Ulgen

                                                                  S.

cerevisiae                                                  YPG/AB
24




                           40

                            4

               4.5

batch                fed-batch

                        batch

        47.5
25




-                  3.1

-          3.2

-                            3

-                        3

-           3.5

-         3.6

-    3.7

-   3.8

-            3.9

-          3.10
26



-              3.11

-         3
-
                3.13

              3.14

                 3.15

              3.16




    3.1
27




3.2
28
29
30



     .   .   .




1.


2.
3.
4.



5.




8.
31




-
32




ml
33




4.1.
34



         40

         40

         20

         40

         6


         6


         20

        2500




2535.




         6
35




        -


        -


        -

        -




    1



1
2
3
4




            4.2
36




  6                                15
                      15       2555 –   30
2555         5                               ml
                           5




                 50


  25




       4.3
37



                 15




             200 ml.



     -

             11.19

70                     54.40



     -

             12.25

80                     1.38



     -

             11.45

70                     50.37



         -
38



          13.21

75                1.17




                         19
ml.
                         16
ml.
                         17.5
ml.
                         20.5
ml.




      2
39




                 ˚c


11.19   54.40    70
12.25   1.38     80
11.45   50.37    70
13.21   1.17     75
40




1.




     2
41



                                6
               15         5




        3

                     200 ml.


-

            11.19              70

                               54.40



    -

             12.25              80

                               1.38



    -
42



          11.45     70

                  50.37




      -

          13.21     75

                   1.17




                          19
ml.
                          16
ml.
                          17.5
ml.
                          20.5
ml.
43




             6
        15


4.2.1




                  40

                  40

                  20

                  40

                  6

                  6


                  20

                 2500
44




2535.
45




      Amylomyces Rhizopus Chlamydomucor




                          Saccharomyces
cerivisiae
fermentation)



lactic acid bacteria         Pediococcus Lactobacillus
                         acetic acid bacteria)
      Acetobacter      Gluconobacter

                                              Amylomyces
Rhizopus



Amylomyces spp.                         0.88-1.29 %
Rhizopus spp.                     3.7-4.3 %
                                            Rose Bengal
dichloran chloramphenical agar (RBDC)         soluble starch 4 %



                         1              0      Rhizopus spp.
                          Amylomyces spp.
46




Saccharomycopsis fibuligera
(                                                                10
              20                                      Saccharomyces
cerevisiae RIT I (                                               S.
fibuligera
                     S. cerevisiae RIT I


                                                           S. fibuligera
             30                       4.0
                                    100
                           10, 20        30
                               S. fibuligera
                                       0. 5, 10        15 %)



                                                       S. fibuligera
20
                                                 S. fibuligera    20
                                 killer strain       sensitive strain

      :                         .2546.


.
47



                                          rDNA                  D1/D2
 domain                      5’          26S
rRNA                ITS-5.8S
rDNA
PCR            D1/D2 domain              600
bp                                 PCR              ITS-5.8S
rDNA
     340-1040 bp        PCR                                   direct
sequencing                                                GenBank
             Blast search
tool                                                    19
                                  Candida atlantica, C.
colliculosa, C. guilliermondii, C. lusitaniae, C.
tropicalis, Hanseniaspora osmophila, H. uvarum, H.
vineae, Loderomyces elongisporus, Pichia anomala,
Saccharomyces cerevisiae            Saccharomycopsis
fibuligera



       API 20C AUX                                           Clustal
X                                                   Kimura 2-
parameter                   phylogenetic tree           neighbor-
joining                                               phylogenetic
tree                                 2
48



                                  ITS-    S
rDNA
             conventional
method




                                                  ITS-   S
rDNA
                        conventional
method
                                         conventional
method


         conventional
method



    :                   . 2548.


.




             Saccharomycopsis fibuligera )
49




 Saccharomyces cerevisiae )




       Saccharomycopsis
        S.cerevisiae
          Saccharomycopsis




            Saccharomyces




        : school of Biological and Molecular Sciences, Oxford

Brookes University, Headington, Oxford OX3 0BP, UK

                          Saccharomyces cerevisiae

glucoamylase              Rhizopus oryzae             Murai et

al. Appl. Environ. Microbiol., 63, 1362-1366, 1997)

                                  , G-1315,


                                                      2.34% (w

/ w)                               5.32% (w / w)

glucoamylase secreting transformants
50




     : Evaluation of the function of arming yeast displaying

glucoamylase on its cell surface by direct fermentation of corn

to ethanol Original Research Article

Journal of Fermentation and Bioengineering, Volume       , Issue

 ,     , Pages     -



Pediococcus Lactobacillus




                               malolactic fermentation    MLF)

           MLF


                          pH
51



       :           .                   8

60 (                   2544)

           4.2.2




                                       1



                                               1   3   1     4


           3

                                   3


                          “    ”




                                   “       ”




                                                           1–2
                   1
52



       :            .           8
60 (                    2544)

            4.2.3




                    50


   25




       -




        -
53



                       10-25




-




-




    PDF


                       amyl alcohol


    amyl alcohol   1
54



     : Kinetics of amyl alcohol production during ethanol

fermentation of blackstrap molasses Original Research Article

Biomass, Volume 1, Issue 2, October 1981, Pages 115-126

W. Borzani, M.L.R. Vairo, L.H. Koshimizu, M.R. De Melo Cruz,

L. Perego Jr.                                        2


                                                 2




2
55



                        100

                 78.3




        70-80


.   .




                70-80




         2.
56




                                               ,


                                      ,


                 .2555.           .       .    :
http://www.surathai.net.
            26             2555
57




              . 2537.                                                                        .

                                                                                    ,

2537.

                    (               .               :
http://web.ku.ac.th/schoolnet/snet5/distill.html 9                                2555


                        (               .               :
http://www.il.mahidol.ac.th 9                       2555



                            (               .                :

http://www.korattreat.net/node/19 9                              2555

                                (               .                 :
http://www.vcharkarn.com/vcafe/60235/2 9                                    2555

                   .2555.                               .             .                 :
http://www.ku.ac.th/e-magazine/january                      agri/kmak/        .         26
         2555




         Zymomonas mobilis,                                               , 2545.
58



                                                               .2555
.
                                .         .          :
    http://library.cmu.ac.th/rsc/?newsdetail.php&id=96.           26
            2555




                                                                    ,
                       ,
                                           5,
                 , 2552.




                                                         ,


                           , 2552.




            Rhizopus sp.             ,              , 2553.
Altntas, M.M., Ulgen, O.K., Kirdar, B. Improvement of ethanol

production      from   starch   by   recombinant    yeast     through

manipulation of environmental factors. Enzyme Microb. Tech.

31: 640-647, 2002.
59



Dung, N.T.P., Rombouts, F.M. Functionality of selected strains

of moulds and yeasts from Vietnamese rice wine starers. Food

Microbiol. 23:331-340, 2006.

Tsuyoshi, N., Fudou, R., Yamanaka, S. Identification of yeast

strains isolated from marcha in Sikkim a microbial starter for

amylolytic fermentation. Int. J. Food Microbiol. 99 : 135 – 146.

2005.

Ulgen, O.K., Saygili, B., Onsan, Z.I. Bioconversion of starch

into ethanol by a recombinant Saccharomyces serevisiae strain

YPG-AB. Process. Biochem. 37 : 1157-1168, 2002
60
61
62
63
64
65
66
67




:

        7                    . .2534   21

                :       72     2

            52150



    -


    -




    -                                       3




            :

        :

                    :
68



                 :




    -

:

        16                . .2534   21

             :       16     4

        54120



    -
69


-


-



-               3




        :

    :

            :

            :

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วิจัยการหมักเอทานอล ครั้ง3

  • 1. 1 1 –––– 1870
  • 2. 2
  • 4. 4 1. (Operational Definition) - 6
  • 5. 5 - - - - -
  • 6. 6 2
  • 7. 7 ( ) 1. 2 - 3
  • 8. 8 2. 3. 4. 5. 6. 2 2-3 6 1.5 – 5.0 2-5 1
  • 9. 9 2.1.1 1. 2. 45%
  • 10. 10 400 6 6 6 6 6 2 2.2 2.2.1 ( ) Amylomyces sp.
  • 11. 11 Aspergillus sp. Rhizopus sp. Mucor Saccharomycopsis sp. Pichia sp. Saccharomyces sp. + + 2.2.2
  • 12. 12 C6H12O6 - - - > 2 C2H5OH + 2CO2 C6H12O6 + 6O2 - - - > 6CO2 + H2O + 2.3 2.3.1 simple distillation) 2.3.2 (fractional distillation) 2.3.3
  • 13. 13 2.3.1 simple distillation) simple distillation) 80 1. 2. boiling stone boiling chips 3. 4.
  • 14. 14 5. 2-3 6. 1. 2. 3. 4. 2.3.2 (fractional distillation) 2
  • 15. 15
  • 16. 16 2.3.3 2.4 2 100
  • 17. 17 78.3 10% 93 55% 82 % 78 78 – 85 90 ( )
  • 18. 18 2.4.1 1. simple distillation) ( )
  • 19. 19 2. (fractional distillation)
  • 20. 20 (2 ) % Saccharomyces cerevisae x / modified Lane-Eynon % HPLC % , % % % % % - Enterobacteraerogenes - (CGWO) 37 oC
  • 21. 21 Conversion ( ) ( 30oC) Conversion CGWO Conversion - - 30 g/l 4 1 ( . ) 2, 3 4 Saccharomyces cerevisiae NP 01, S. bayanus EC 1118 S. cerevisiae Sweden 2 35 32 10 2 - 500
  • 22. 22 5 Z. mobilis 5 Z. mobilis TISTR 548 100 - (pH) 5.5 100 0.463 NH4Cl 1 0.419 Altintas Saccharomyces cerevisae YPB-G 4
  • 23. 23 0.118 0.183 Dung 29 119 53 15 51 Amylomy ces rouxii Saccharomyces cerevisiae Tsuyoshi 6 20 4 1 Saccharomyces bayanus 2 Candida glabrata 3 Pichia anomala 4 Saccharomycopsis fibuligera 1,2 3 Ulgen S. cerevisiae YPG/AB
  • 24. 24 40 4 4.5 batch fed-batch batch 47.5
  • 25. 25 - 3.1 - 3.2 - 3 - 3 - 3.5 - 3.6 - 3.7 - 3.8 - 3.9 - 3.10
  • 26. 26 - 3.11 - 3 - 3.13 3.14 3.15 3.16 3.1
  • 28. 28
  • 29. 29
  • 30. 30 . . . 1. 2. 3. 4. 5. 8.
  • 31. 31 -
  • 32. 32 ml
  • 34. 34 40 40 20 40 6 6 20 2500 2535. 6
  • 35. 35 - - - - 1 1 2 3 4 4.2
  • 36. 36 6 15 15 2555 – 30 2555 5 ml 5 50 25 4.3
  • 37. 37 15 200 ml. - 11.19 70 54.40 - 12.25 80 1.38 - 11.45 70 50.37 -
  • 38. 38 13.21 75 1.17 19 ml. 16 ml. 17.5 ml. 20.5 ml. 2
  • 39. 39 ˚c 11.19 54.40 70 12.25 1.38 80 11.45 50.37 70 13.21 1.17 75
  • 40. 40 1. 2
  • 41. 41 6 15 5 3 200 ml. - 11.19 70 54.40 - 12.25 80 1.38 -
  • 42. 42 11.45 70 50.37 - 13.21 75 1.17 19 ml. 16 ml. 17.5 ml. 20.5 ml.
  • 43. 43 6 15 4.2.1 40 40 20 40 6 6 20 2500
  • 45. 45 Amylomyces Rhizopus Chlamydomucor Saccharomyces cerivisiae fermentation) lactic acid bacteria Pediococcus Lactobacillus acetic acid bacteria) Acetobacter Gluconobacter Amylomyces Rhizopus Amylomyces spp. 0.88-1.29 % Rhizopus spp. 3.7-4.3 % Rose Bengal dichloran chloramphenical agar (RBDC) soluble starch 4 % 1 0 Rhizopus spp. Amylomyces spp.
  • 46. 46 Saccharomycopsis fibuligera ( 10 20 Saccharomyces cerevisiae RIT I ( S. fibuligera S. cerevisiae RIT I S. fibuligera 30 4.0 100 10, 20 30 S. fibuligera 0. 5, 10 15 %) S. fibuligera 20 S. fibuligera 20 killer strain sensitive strain : .2546. .
  • 47. 47 rDNA D1/D2 domain 5’ 26S rRNA ITS-5.8S rDNA PCR D1/D2 domain 600 bp PCR ITS-5.8S rDNA 340-1040 bp PCR direct sequencing GenBank Blast search tool 19 Candida atlantica, C. colliculosa, C. guilliermondii, C. lusitaniae, C. tropicalis, Hanseniaspora osmophila, H. uvarum, H. vineae, Loderomyces elongisporus, Pichia anomala, Saccharomyces cerevisiae Saccharomycopsis fibuligera API 20C AUX Clustal X Kimura 2- parameter phylogenetic tree neighbor- joining phylogenetic tree 2
  • 48. 48 ITS- S rDNA conventional method ITS- S rDNA conventional method conventional method conventional method : . 2548. . Saccharomycopsis fibuligera )
  • 49. 49 Saccharomyces cerevisiae ) Saccharomycopsis S.cerevisiae Saccharomycopsis Saccharomyces : school of Biological and Molecular Sciences, Oxford Brookes University, Headington, Oxford OX3 0BP, UK Saccharomyces cerevisiae glucoamylase Rhizopus oryzae Murai et al. Appl. Environ. Microbiol., 63, 1362-1366, 1997) , G-1315, 2.34% (w / w) 5.32% (w / w) glucoamylase secreting transformants
  • 50. 50 : Evaluation of the function of arming yeast displaying glucoamylase on its cell surface by direct fermentation of corn to ethanol Original Research Article Journal of Fermentation and Bioengineering, Volume , Issue , , Pages - Pediococcus Lactobacillus malolactic fermentation MLF) MLF pH
  • 51. 51 : . 8 60 ( 2544) 4.2.2 1 1 3 1 4 3 3 “ ” “ ” 1–2 1
  • 52. 52 : . 8 60 ( 2544) 4.2.3 50 25 - -
  • 53. 53 10-25 - - PDF amyl alcohol amyl alcohol 1
  • 54. 54 : Kinetics of amyl alcohol production during ethanol fermentation of blackstrap molasses Original Research Article Biomass, Volume 1, Issue 2, October 1981, Pages 115-126 W. Borzani, M.L.R. Vairo, L.H. Koshimizu, M.R. De Melo Cruz, L. Perego Jr. 2 2 2
  • 55. 55 100 78.3 70-80 . . 70-80 2.
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