非甲醇诱导重组毕赤酵母表达木聚糖酶的条件优化

蓝娜娜 , 张薷月 , 苏然 , 等

doi:10.13995/j.cnki.11-1802/ts.015358

食品与发酵工业 >

2018 , Vol. 44 >Issue 3: 8

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.015358

非甲醇诱导重组毕赤酵母表达木聚糖酶的条件优化

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(上海理工大学医疗器械与食品学院，食品科学与工程研究所，上海，200093)

硕士研究生

网络出版日期: 2018-03-26

基金资助

国家自然科学基金资助项目(21306112)；上海市自然基金(13ZR1429100)；教育部留学回国人员科研启动基金

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Optimization of xylanase expression by recombinant Komagataella phaffii without methanol

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(Institute of Food Science and Engineering, School of Medical Instrument and Food Engineering, University of Shanghai for Science and Technology, Shanghai 200093, China)

Online published: 2018-03-26

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摘要

以1株表达木聚糖酶的重组毕赤酵母为试材，首先优化了甲醇诱导的培养条件，进而采用甲酸铵作为诱导剂，联合山梨醇或者甲醇诱导重组毕赤酵母表达木聚糖酶。结果表明，甲醇诱导时木聚糖酶活力最高达到3 403.8 U/mL，单位细胞浓度活力为97.5 U/mL OD600。甲酸铵诱导毕赤酵母醇氧化酶1启动子的强度为甲醇诱导时的40.9%。联合0.5%甲酸铵和0.5%山梨醇时木聚糖酶活力为2 032.0 U/mL，单位细胞浓度的木聚糖酶活力为甲醇诱导时的1.54倍。这表明甲酸铵可以作为非甲醇诱导剂诱导毕赤酵母表达外源蛋白，并且联合山梨醇可以使重组毕赤酵母高效表达外源蛋白。

关键词： 重组毕赤酵母; color:#2453B3; font-family:"; ????"; font-size:12px; font-style:normal; font-weight:700; line-height:18px; text-decoration:none; ">; 木聚糖酶; color:#2453B3; font-family:"; ????"; font-size:12px; font-style:normal; font-weight:700; line-height:18px; text-decoration:none; ">; 甲酸铵; color:#2453B3; font-family:"; ????"; font-size:12px; font-style:normal; font-weight:700; line-height:18px; text-decoration:none; ">; 混合碳源诱导

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蓝娜娜 , 张薷月 , 苏然 , 等 . 非甲醇诱导重组毕赤酵母表达木聚糖酶的条件优化[J]. 食品与发酵工业, 2018 , 44(3) : 8 . DOI: 10.13995/j.cnki.11-1802/ts.015358

Abstract

Komagataella phaffii is a famous host for heterologous protein expression. However, methanol is needed as inducer and lead to toxic and explosive danger when it is used at large amount of industrial scale. Therefore, non-methanol inducer is urgent needed and thus become a hot topic in this research field. In this research, a recombinant Komagataella phaffii expressing xylanase was used to explore non-methanol inducer. The conditions of methanol induction were optimized to obtain 3 403.8 U/mL xylanase, and 97.5 U/mL OD600 specific xylanase. 40.9% intensity of AOX1 promoter under methanol induction was obtained under 0.5% ammounim formate induction. And 2 032.0 U/mL xylanase was obtained at the condition of 0.5% ammonium formate and 0.5% sorbitol addition per 24 h, which was 1.54 fold of AOX1 promoter intensity at the condition of methanol induction. Therefore, ammonium formate combined with sorbitol was an efficient inducer for heterologous protein expression of Komagataella phaffii.

Key words： recombinant Komagataella phaffii; color:#2453B3; font-family:"; ????"; font-size:12px; font-style:normal; font-weight:700; line-height:18px; text-decoration:none; ">; xylanase; color:#2453B3; font-family:"; ????"; font-size:12px; font-style:normal; font-weight:700; line-height:18px; text-decoration:none; ">; ammonium formate; color:#2453B3; font-family:"; ????"; font-size:12px; font-style:normal; font-weight:700; line-height:18px; text-decoration:none; ">; mix-carbon induction

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