文献类型：期刊文献

中文题名：适应性进化与甜菜碱的添加促进安丝菌素的生产

英文题名：Adaptive evolution and addition of betaine to promote ansamitocin production

作者：许文龙[1];胡凤仙[1];刘婷[1];韦柳静[1];花强[1]

机构：[1]华东理工大学生物工程学院生物反应器工程国家重点实验室

年份：2019

卷号：46

期号：7

起止页码：1561

中文期刊名：微生物学通报

外文期刊名：Microbiology China

收录：CSTPCD;;北大核心:【北大核心2017】；CSCD:【CSCD2019_2020】；

基金：高等学校博士学科点专项科研基金(20130074110002)~~

语种：中文

中文关键词：适应性进化;珍贵橙色束丝放线菌;安丝菌素P-3;甜菜碱

外文关键词：Adaptive evolution;Actinosynnema pretiosum;ansamitocin P-3;betaine

摘要：【背景】安丝菌素是一类由珍贵束丝放线菌橙色亚种(Actinosynnema pretiosum ssp.auranticu)生产的美登素类衍生抗生素,属于大环内酰胺类物质,根据不同的C-3位基团可以分为一系列衍生物。目前已上市的高效抗癌药物曲妥珠单抗(T-DM1)以AP-3 (Ansamitocin P-3)为生产底物,并表现出很好的乳腺癌治疗效果。然而当前AP-3的产量较低,其过高的成本限制了进一步发展。【目的】应用适应性进化及添加适量甜菜碱策略提高安丝菌素AP-3的产量。【方法】以珍贵束丝放线菌橙色亚种为出发菌株,链霉素和巴龙霉素为胁迫压力进行适应性进化,筛选出安丝菌素积累较高的菌株,随后向进化菌株的发酵培养基中添加0.1%的甜菜碱,AP-3的产量进一步提高,同时分析了进化菌株AP-3相关基因的转录水平,初步探索进化菌株安丝菌素积累提高的原因。【结果】得到2株进化菌株Str16-4-4和Par16-2-1,发酵7d后AP-3产量分别提高了33.4%和31.7%,添加甜菜碱后其AP-3产量相比出发菌株提高了54.6%和47.4%。【结论】通过适应性进化的策略获得了AP-3生产能力提高的珍贵束丝放线菌橙色亚种进化菌株,对促进AP-3的生产提供了新思路,而且为适应性进化策略提高目标产物的产量提供了新的例证。
[Background] Ansamitocins,macrocyclic lactam compounds related to maytansine,are synthesized by Actinosynnema pretiosum ssp.auranticun.Based on the differences of ester side chain moieties in C3 position,ansamitocins contain a series of derivatives.Currently,a novel anticancer drug named Kadcyla(T-DM1),formed from ansamitocin P-3(AP-3),enters the market.T-DM1 had been approved by FDA as an antibody-drug for treating breast cancer.However,the low yield of ansamitocins limits its commercial application.[Objective] To improve AP-3 yield from A.pretiosum ssp.auranticum ATCC 31565,we applied adaptive evolution and addition of betaine.[Methods] The wild-type A.pretiosum ssp.auranticum was used as starting strain to the adverse stress streptomycin and paromomycin.The evolved mutants were screened with better production of AP-3.After 0.1% betaine was added into the fermentation broth of the evolved mutants,the production of AP-3 was further enhanced.Then,gene transcription levels related to AP-3 biosynthesis were analyzed in evolved mutants and starting strain,to investigate the reasons for enhancement of ansamitocin.[Results] Two evolved strains Str16-4-4 and Par16-2-1 have been obtained.After 7 days fermentation,their AP-3 contents were increased by 33.4% and 31.7%,respectively.Addition of0.1% betaine ultimately resulted in an increase of 54.6% and 47.4% AP-3 production compared to the wild strain.[Conclusion] Through the adaptive evolution strategy,AP-3 production was improved in A.pretiosum ssp.auranticum.This study provides a new strategy to improve the production of AP-3,and also gives a new example for adaptive evolutionary strategy to promote the production of target products.