苯基乳酸耐受性大肠杆菌的筛选及其高产苯基乳酸特性

doi:10.7506/spkx1002-6630-201714004

食品科学 ›› 2017, Vol. 38 ›› Issue (14): 24-29.doi: 10.7506/spkx1002-6630-201714004

苯基乳酸耐受性大肠杆菌的筛选及其高产苯基乳酸特性

徐艳，郭倩，朱益波，王立梅，齐斌

（1.苏州大学基础医学与生物科学学院，江苏?苏州 215000；2.常熟理工学院生物与食品工程学院，苏州市食品生物技术重点实验室，发酵工程技术研究中心，江苏?常熟 215500）

出版日期:2017-07-25 发布日期:2017-09-06
基金资助:
国家自然科学基金面上项目（31470092）；国家自然科学青年科学基金项目（31501459）；江苏省“六大高峰人才”资助计划项目（NY-021）

Screening of Phenyllactic Acid-Resistant Escherichia coli and Its Application in the Synthesis of Phenyllactic Acid

XU Yan, GUO Qian, ZHU Yibo, WANG Limei, QI Bin

(1. School of Biology & Basic Medical Sciences, Soochow University, Suzhou 215000, China; 2. Key Laboratory of Food and Biotechnology of Suzhou, Fermentation Engineering Technology Research Center, College of Biological and Food Engineering, Changshu Institute of Technology, Changshu 215500, China)

Online:2017-07-25 Published:2017-09-06

摘要/Abstract

摘要： 苯基乳酸（phenyllactic acid，PLA）耐受性菌株的筛选能够有效降低PLA对生产菌株的抑制作用，有利于PLA产量的提高。通过紫外诱变的方法筛选耐受PLA的菌株，并应用于PLA的合成。以Escherichia coli BL21（DE3）作为原始出发菌株，通过紫外诱变的方法诱变筛选获得一株耐受PLA突变菌株E. coli Z2016（CCTCC保藏编号M2016332）。以E. coli Z2016为宿主菌，分别构建了重组菌株E. coli Z2016 pET-28a-ldhY52V和E. coli Z2016 pET-28a-ldhL用于D-和L-苯基乳酸的合成。结果表明：E. coli Z2016在含有1 g/L D-PLA的培养基中能够正常生长；重组突变菌株E. coli Z2016 pET-28a-ldhY52V和E. coli Z2016 pET-28a-ldhL全细胞合成D-PLA和L-PLA产量分别为6.75、6.97 g/（L·h），较重组出发菌株分别提高了14.02%和8.95%；分批补加底物反应120 min，E. coli Z2016 pET-28a-ldhY52V得到的D-PLA为20.02 g/L，较对照组提高22.17%，转化率为90.07%；E. coli Z2016 pET-28a-ldhL得到的L-PLA产量为20.87 g/L，较对照组提高16.85%，最终转化率为91.24%。筛选耐受性菌株是提高PLA产量的有效途径。

关键词: 紫外诱变, 苯基乳酸, 苯基乳酸耐受菌株, 大肠杆菌, 全细胞转化

Abstract: Screening phenyllactic acid (PLA)-resistant strains enables effective reduction of the inhibitory effect of PLA on the synthesis process and thus an increase in PLA production. In this report, Escherichia coli BL21 (DE3) was mutagenized by UV irradiation and screened for a mutant resistant to PLA, E. coli Z2016 (CCTCC Accession No. M2016332). E. coli Z2016 was used the host strain to construct recombinant strains E. coli Z2016 pET-28a-ldhY52V and E. coli Z2016 pET-28a-ldhL for the synthesis of D- and L-PLA. The results showed that E. coli Z2016 was able to grow normally in a medium containing 1 g/L D-PLA. The production of D- and L-PLA by the recombinant strains were 6.75 and 6.97 g/(L·h), which were increased by 14.02% and 8.95% as compared to those produced by the original strain, respectively. During 120 min of fed-batch fermentation, the production of D-PLA by E. coli Z2016 pET-28a-ldhY52V was 20.02 g/L with a conversion rate of 90.97%, which was increased by 22.17% as compared to that produced by the control. The L-PLA concentration produced by E. coli Z2016 pET-28a-ldhL was 20.87 g/L, which was increased by 16.85% as compared to produced by the control, with a conversion rate of 91.24%. Accordingly, the use of PLA-resistant strains provided an effective method to increase PLA production.

Key words: UV mutagenesis, phenyllactic acid, phenyllactic acid-resistant strain, E. coli, whole-cell transformation

中图分类号:

TS201.2

徐艳，郭倩，朱益波，王立梅，齐斌. 苯基乳酸耐受性大肠杆菌的筛选及其高产苯基乳酸特性[J]. 食品科学, 2017, 38(14): 24-29.

XU Yan, GUO Qian, ZHU Yibo, WANG Limei, QI Bin. Screening of Phenyllactic Acid-Resistant Escherichia coli and Its Application in the Synthesis of Phenyllactic Acid[J]. FOOD SCIENCE, 2017, 38(14): 24-29.

[1]	甘玉飞，薛正莲，周杰，王芳，王洲，刘艳. 黏质沙雷菌磷脂酶A1辅助蛋白S对表达宿主菌大肠杆菌抑制作用机制[J]. 食品科学, 2021, 42(7): 45-51.
[2]	门佳轩，熊博，郝亚男，李旋，刘益宁，谢希贤. 代谢工程优化大肠杆菌高效合成L-苯丙氨酸[J]. 食品科学, 2021, 42(2): 114-120.
[3]	孙悦，刘佳伊，陈璐，杜宏，白凤翎，吕欣然，张德福，郭晓华，励建荣. 抗耐药性大肠杆菌乳酸菌的筛选及抑菌机制[J]. 食品科学, 2021, 42(2): 121-127.
[4]	王虹懿，刘芳，孙芝兰，苏萌萌，诸永志，王道营，徐为民，彭景. Helveticin-M与绿原酸复配对大肠杆菌和肠炎沙门氏菌的抑菌效果及其机制[J]. 食品科学, 2020, 41(3): 68-74.
[5]	吴丽娜，董鹏程，张一敏，毛衍伟，梁荣蓉，杨啸吟，朱立贤，罗欣. 大肠杆菌O157:H7在不锈钢表面生物膜形成能力及其与菌株的特性关系[J]. 食品科学, 2020, 41(22): 127-132.
[6]	李强，韩亚昆，蒋帅，张悦，吴鹤云，徐庆阳，谢希贤. 代谢工程改造大肠杆菌合成反式-4-羟基-L-脯氨酸[J]. 食品科学, 2020, 41(2): 202-207.
[7]	夏丹丹，赵莹莹，马盼盼，王深垒，马常阳，郜晓峰，康文艺. 大肠杆菌DNA定性标准物质的制备及在枸杞子污染检测中的应用[J]. 食品科学, 2020, 41(18): 267-274.
[8]	余兰林，姬赛赛，禹金龙，付文静，张林，李蛟龙，高峰，江芸. 盐应激对大肠杆菌O157:H7存活和毒力基因表达的影响[J]. 食品科学, 2020, 41(14): 95-101.
[9]	吴克刚，赵欣欣，段雪娟，柴向华，于泓鹏，刘晓丽，范宇婷. 芳樟醇气相抗菌活性与作用机制[J]. 食品科学, 2020, 41(1): 61-67.
[10]	李萍，林洪，童贻刚，王静雪. 1株T1样噬菌体vB_EcoS_IME18的分离鉴定及其受体分析[J]. 食品科学, 2019, 40(8): 79-86.
[11]	禹金龙，董晨，王娴静，姬赛赛，付文静，胡洁，江芸. 牛源性非O157大肠杆菌的分离与鉴定[J]. 食品科学, 2019, 40(4): 299-304.
[12]	张爱静，李琳琼，朱蕾，王鹏杰，高瑀珑. 热胁迫和培养温度对大肠杆菌抗热性的影响[J]. 食品科学, 2019, 40(22): 75-80.
[13]	周陆红，张鹏飞，张杰，吴聪明，唐晓双，郝丹，王新. 屠宰猪中大肠杆菌毒力基因检测及耐药性分析[J]. 食品科学, 2019, 40(2): 264-268.
[14]	王娴静，董晨，禹金龙，胡洁，付文静，张苏，江芸. 酸应激对大肠杆菌O157:H7生物菌膜形成的影响[J]. 食品科学, 2019, 40(19): 1-7.
[15]	王淑娟，刘程，方水琴，田亚晨，董庆利，王翔，许东坡，刘箐,. 基于钴金属有机骨架材料（ZIF-67）催化性能的牛奶中大肠杆菌O157:H7快速定量检测[J]. 食品科学, 2019, 40(18): 323-328.

苯基乳酸耐受性大肠杆菌的筛选及其高产苯基乳酸特性

Screening of Phenyllactic Acid-Resistant Escherichia coli and Its Application in the Synthesis of Phenyllactic Acid

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价