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- Title
霍氏肠杆菌β-胡萝卜素9,10'双加氧酶的制备及其应用.
- Authors
魏涛; 王敏; 张志平; 段乃心; 杨旭; 黄申; 毛多斌
- Abstract
Using the genome DNA of Enterobacter honnaechei as template, the gene of β-carotene-9,10'-dioxygenase was amplified by polymerase chain reaction (PCR), the recombinant plasmid was constructed to express in Escherichia coli BL21(DE3), and the enzyme activity was detected by HPLC. The results showed that the purified recombinant β-carotene 9,10' dioxygenase was obtained by Nichelating affinity chromatography and molecular sieve Sephacryl TM 5-200. The results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that the molecular mass of the enzyme was approximately 57 kDa, and the optimum reaction temperature and pH of the enzyme were 40 t and 8.5, respectively. When the substrate β-carotene mass concentration was 500 mg/L and β-carotene 9,10' dioxygenase activity was 0.4 U/ml, the β-ionone yield was 142.3 mg/L and the yield reached 79.4%.
- Subjects
MOLECULAR weights; AFFINITY chromatography; POLYMERASE chain reaction; MOLECULAR sieves; GEL electrophoresis
- Publication
China Brewing, 2022, Vol 41, Issue 1, p128
- ISSN
0254-5071
- Publication type
Article
- DOI
10.11882/j.issn.0254-5071.2022.01.022