重组毕赤酵母中高水平分泌及环糊精葡萄糖基转移酶的特性研究。
High-level secretion and characterization of cyclodextrin glycosyltransferase in recombinant Komagataella phaffii.
机构信息
Institute of Food Science and Engineering, University of Shanghai for Science and Technology, Shanghai 200093, PR China.
Institute of Food Science and Engineering, University of Shanghai for Science and Technology, Shanghai 200093, PR China.
出版信息
J Biotechnol. 2017 Oct 10;259:126-134. doi: 10.1016/j.jbiotec.2017.07.031. Epub 2017 Jul 28.
Cyclodextrin glycosyltransferase (CGTase) catalyzes the conversion of starch into cyclodextrin (CD), which is widely applied in food, pharmaceutical, cosmetic, and agricultural industries. For efficient production of CD, high yield of CGTase with good characteristics is necessary. In this study, the cgt gene from Bacillus pseudalcaliphilus was expressed in Komagataella phaffii after codon optimization and expression vector selection. The β-CGTase activity in the transformant reached 3885.1UmL, which is the highest value reported so far, at 28°C, 6% inoculum ratio, and 1.5% methanol addition following 24h of incubation. The recombinant CGTase showed high specific activity at 80°C without any γ-CGTase activity, and had good stability in a wide pH and temperature range. These results demonstrate that the recombinant CGTase could have potential industrial applications.
环糊精葡萄糖基转移酶（CGTase）可催化淀粉转化为环糊精（CD），CD 在食品、制药、化妆品和农业等行业有广泛应用。为了高效生产 CD，需要高产、特性良好的 CGTase。本研究对来自解淀粉芽孢杆菌的 cgt 基因进行密码子优化和表达载体选择，在毕赤酵母中表达。在 28°C、6%接种量、1.5%甲醇添加、24h 发酵条件下，转化体的β-CGTase 酶活达到 3885.1UmL，是迄今为止报道的最高值。重组 CGTase 在 80°C 时具有高比活，且无 γ-CGTase 活性，在较宽的 pH 和温度范围内具有良好的稳定性。这些结果表明，重组 CGTase 具有潜在的工业应用价值。
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