发酵法生产利普司他汀的研究进展

doi:10.16085/j.issn.1000-6613.2020-1653

摘要/Abstract

摘要：

利普司他汀（lipstatin）是一种β-内酯分子，能选择性抑制胰脂肪酶的活性，从而减少小肠脂肪的吸收。分析表明，一个利普司他汀分子与一个脂肪酶分子结合，通过活性部位丝氨酸残基的酰化来抑制胰脂肪酶的催化活性。利普司他汀由毒三链霉菌（Streptomyces toxytricini，简称S. toxytricini）生产的一种天然产物。本文综述了利普司他汀的结构特点，分析了利普司他汀市场应用现状，着重论述了发酵法生产利普司他汀的研究进展；提出生物合成利普司他汀的机制尚不清楚，但酰基辅酶A羧化酶（ACCase）复合物在利普司他汀生物合成中起着关键作用。最后，指出当前存在的问题并对代谢工程在利普司他汀产生菌发酵中的应用进行概述，对今后的研究趋势进行了展望。

关键词: 利普司他汀, 发酵, 代谢工程, 毒三素链霉菌

Abstract:

Lipstatin is a β-lactone molecule which controls the digestive activity of pancreatic lipases and thus controls the absorption of fat in the small intestine. Lipstatin is supposed to inhibit the catalytic activity of pancreatic lipase by acylation of the serine residue present in the active site. A binding study suggests that one molecule of lipstatin binds with one molecule of lipase. Lipstatin is a natural product produced from Streptomyces toxytricini. The chemical structure is introduced,and the current application situation is reviewed. The progress in production of lipstatin fermentation is presented with a discussion on existing problems. Technology of metabolic engineering used in fermentation is introduced especially.The mechanism of lipstatin biosynthesis in bacteria is not yet clear, however acyl-coenzyme A carboxylase (ACCase) complex is playing key role in lipstatin biosynthesis. Furthermore, prospect for future research is proposed.

Key words: lipstatin, fermentation, metabolic engineering, Streptomyces toxytricini

中图分类号:

TQ460

李玲, 于泳, 胡永红. 发酵法生产利普司他汀的研究进展[J]. 化工进展, 2021, 40(4): 2251-2257.

LI Ling, YU Yong, HU Yonghong. Research progress in production of lipstatinfermentation[J]. Chemical Industry and Engineering Progress, 2021, 40(4): 2251-2257.

图/表 4

参考文献 48

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菌株	育种、优化方法	效价 /mg·L^-1	研究团队	年份
S.toxytricini NRRL 15443		6.23	Paul等^[22]	1985
S.toxytricini NRRL 15443		43.17	Weibel等^[2]	1987
S.toxytricini CCM 7349	70L发酵罐补料发酵，约34h补加亚油酸、L-亮氨酸混合溶液	3290	Varga等^[23]	2007
S.toxytricini	补料添加0.3%亚油酸、1%豆油	4773	Gasparic等^[24]	2007
S.toxytricini	培养20h后补加亚油酸和L-亮氨酸，控制亚油酸体积分数0.3%~0.5%	4400	Fujs等^[25]	2010
S.toxytricini NRRL 15443	Plackett-Burman法优化发酵种子、发酵培养基	1980	Kumar等^[26]	2011
S.toxytricini ATCC 19813	UV和NTG联合诱变，全因子试验设计优化发酵培养条件，大豆粉35g/L和豆油25g/L	3290	Luthra等^[27-28]	2013
S.toxytricini NRRL B-5426	调节碳氮比例、补充金属辅因子、维生素、TCA循环中间产物，有机酸和NADH	2242.63	Kumar等^[29]	2016
S.toxytricini NRRL B-5426	紫外、甲磺酸乙酯、甲基甲烷磺酸盐和NTG复合诱变结合前体物添加	2387	Kumar等^[31]	2018
S.toxytricini srn7	添加前体：亚油酸0.02～0.1mg/g，油酸0.02～0.15mg/g，L-亮氨酸1.5%，大豆粉10%	5886	Khushboo等^[32]	2020
S.toxytricini HCCB-2	紫外和微波复合诱变，培养条件优化	663	王栋海^[33]	2004
S.toxytricini XC-lp-69	紫外和微波复合诱变，补料发酵	1400	韩俊如等^[34]	2006
S.toxytricini SIPI-UN-21	紫外、NTG复合诱变，培养条件优化	3.25倍	黄娟等^[38]	2006
S.toxytricini	正交设计优化发酵培养基，补料油酸/辛酸补料、L-亮氨酸和豆油	1.2倍	王永青^[36]	2013
S.toxytricini zjut011	正交试验法，添加前体L-亮氨酸，首次发现二价阳离子Mg²⁺、Co²⁺和Zn²⁺对提高效价有显著作用	4208	Zhu等^[35]	2014
S.toxytricini FIM-1015	考察溶氧和剪切力因素，10L发酵罐补料发酵	6446	孙菲^[37]	2014
S.toxytricini LIPS-352	紫外-微波复合诱变，添加前体葵花籽油，Box-Behnken设计优化	8122.9	刘顺娜等^[39]	2017
S.toxytricini FIM-17-16	发酵条件优化	9667	陈洲琴等^[40]	2020

菌株	育种、优化方法	效价 /mg·L^-1	研究团队	年份
S.toxytricini NRRL 15443		6.23	Paul等^[22]	1985
S.toxytricini NRRL 15443		43.17	Weibel等^[2]	1987
S.toxytricini CCM 7349	70L发酵罐补料发酵，约34h补加亚油酸、L-亮氨酸混合溶液	3290	Varga等^[23]	2007
S.toxytricini	补料添加0.3%亚油酸、1%豆油	4773	Gasparic等^[24]	2007
S.toxytricini	培养20h后补加亚油酸和L-亮氨酸，控制亚油酸体积分数0.3%~0.5%	4400	Fujs等^[25]	2010
S.toxytricini NRRL 15443	Plackett-Burman法优化发酵种子、发酵培养基	1980	Kumar等^[26]	2011
S.toxytricini ATCC 19813	UV和NTG联合诱变，全因子试验设计优化发酵培养条件，大豆粉35g/L和豆油25g/L	3290	Luthra等^[27-28]	2013
S.toxytricini NRRL B-5426	调节碳氮比例、补充金属辅因子、维生素、TCA循环中间产物，有机酸和NADH	2242.63	Kumar等^[29]	2016
S.toxytricini NRRL B-5426	紫外、甲磺酸乙酯、甲基甲烷磺酸盐和NTG复合诱变结合前体物添加	2387	Kumar等^[31]	2018
S.toxytricini srn7	添加前体：亚油酸0.02～0.1mg/g，油酸0.02～0.15mg/g，L-亮氨酸1.5%，大豆粉10%	5886	Khushboo等^[32]	2020
S.toxytricini HCCB-2	紫外和微波复合诱变，培养条件优化	663	王栋海^[33]	2004
S.toxytricini XC-lp-69	紫外和微波复合诱变，补料发酵	1400	韩俊如等^[34]	2006
S.toxytricini SIPI-UN-21	紫外、NTG复合诱变，培养条件优化	3.25倍	黄娟等^[38]	2006
S.toxytricini	正交设计优化发酵培养基，补料油酸/辛酸补料、L-亮氨酸和豆油	1.2倍	王永青^[36]	2013
S.toxytricini zjut011	正交试验法，添加前体L-亮氨酸，首次发现二价阳离子Mg²⁺、Co²⁺和Zn²⁺对提高效价有显著作用	4208	Zhu等^[35]	2014
S.toxytricini FIM-1015	考察溶氧和剪切力因素，10L发酵罐补料发酵	6446	孙菲^[37]	2014
S.toxytricini LIPS-352	紫外-微波复合诱变，添加前体葵花籽油，Box-Behnken设计优化	8122.9	刘顺娜等^[39]	2017
S.toxytricini FIM-17-16	发酵条件优化	9667	陈洲琴等^[40]	2020

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