Expressing endoglucanase and α- amylase in Lactobacillus casei E40
碩士 === 大同大學 === 生物工程學系(所) === 106 === Endoglucanase and α-amylase are common industrial enzyme. In this study, the expressing vector pUL6erm-gad-usp-eglB and pUL6erm-gad-usp-amyA, were constructed based on the shuttle vector pUL6erm carrying amylase gene or endoglucanase gene with a chloride-inducib...
| Main Authors: | , |
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| Format: | Others |
| Language: | zh-TW |
| Published: |
2018
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| Online Access: | http://ndltd.ncl.edu.tw/handle/ew92wp |
| Summary: | 碩士 === 大同大學 === 生物工程學系(所) === 106 === Endoglucanase and α-amylase are common industrial enzyme. In this study, the expressing vector pUL6erm-gad-usp-eglB and pUL6erm-gad-usp-amyA, were constructed based on the shuttle vector pUL6erm carrying amylase gene or endoglucanase gene with a chloride-inducible (gadR) expression cassette using Pgad as promoter and could be transformed easily and maintained stably in E. coli, and Lactobacillus casei. The endoglucanase and α-amylase could be induced and expressed in the presence of 0.3 M sodium chloride and 50 mM glutamate. The recombinant endoglucanase was purified with DEAE anion exchange column. Its’ the specific activity was 0.102 U/mg of the partially purified enzyme, the recovery was 43.5% and purified fold was 2.16. The optimal temperature and pH was 37°C and pH 6. The optimal temperature and pH of the crude α-amylase was 50°C and pH 6. Using exo-cellular concentrate of endoglucanase and α-amylase for solid media which contain carboxymethylcelluose or soluble starch assay both showed positive results. This revealed that the transformant strain (Lactobacillus casei E40-10 or Lactobacillus casei E40-A6) can secrete out the recombinant endoglucanase or alpha-amylase into the culture broth, respectively.
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