|本期目录/Table of Contents|

[1]谢慧,马钦元,魏东芝,等.高产柠檬酸工业Aspergillus niger菌株转化体系构建及衣康酸发酵[J].南京工业大学学报(自然科学版),2018,40(06):40-50.[doi:10.3969/j.issn.1671-7627.2018.06.007]
 XIE Hui,MA Qinyuan,WEI Dongzhi,et al.Establishment of transformation system of an industrial Aspergillus niger strain with high citric-acid productivity and itaconic acid fermentation[J].Journal of NANJING TECH UNIVERSITY(NATURAL SCIENCE EDITION),2018,40(06):40-50.[doi:10.3969/j.issn.1671-7627.2018.06.007]
点击复制

高产柠檬酸工业Aspergillus niger菌株转化体系构建及衣康酸发酵()
分享到:

《南京工业大学学报(自然科学版)》[ISSN:1671-7627/CN:32-1670/N]

卷:
40
期数:
2018年06期
页码:
40-50
栏目:
出版日期:
2018-11-20

文章信息/Info

Title:
Establishment of transformation system of an industrial Aspergillus niger strain with high citric-acid productivity and itaconic acid fermentation
文章编号:
1671-7627(2018)06-0040-11
作者:
谢慧12马钦元3魏东芝1王风清1
1.华东理工大学 鲁华生物技术研究所 生物反应器国家重点实验室,上海 200237; 2.河南农业大学 生命科学学院,河南 郑州 450002; 3.潍坊英轩实业有限公司,山东 潍坊 262499
Author(s):
XIE Hui12MA Qinyuan3WEI Dongzhi1WANG Fengqing1
1.State key Laboratory of Bioreactor Engineering,Newworld Institute of Biotechnology,East China University of Science and Technology,Shanghai 200237,China; 2.Life Science College,Henan Agricultural University,Zhengzhou 450002,China; 3.Weifang Ensign Industry Co. Ltd.,Weifang 262499,China
关键词:
柠檬酸 A.niger 农杆菌介导转化 衣康酸
Keywords:
citric acid Aspergillus niger Agrobacterium tumefacien mediated transformation itaconic acid
分类号:
Q815
DOI:
10.3969/j.issn.1671-7627.2018.06.007
文献标志码:
A
摘要:
柠檬酸高产菌株Aspergillus niger YX-1217无法成功建立遗传转化体系,极大地限制了其代谢机制和分子机制的研究。对该菌株进行遗传改造,最终建立了一种农杆菌(Agrobacterium tumefacien)介导的遗传转化体系:农杆菌AGL-1作为介导菌株、A.niger YX-1217孢子密度为 1×107 mL-1、孢子与农杆菌的体积比为1:9、乙酰丁香酮浓度为300 μmol/L。在最佳转化体系下获得25±1个转化子(以107个分生孢子计)。基于上述遗传转化技术,对A.niger YX-1217产柠檬酸的应用进行拓展,通过引入乌头酸脱羧酶cadA基因构建衣康酸的合成途径,基于搅拌转速三阶段控制利用补料分批发酵生产衣康酸,质量浓度最高达到4.21 g/L,最大产率为0.040 g/(L·h),从而延伸原有柠檬酸的代谢通路来实现衣康酸的生产。
Abstract:
Compared with other filamentous fungi,the genetic transformation of Aspergillus niger YX-1217 with high citric-acid productivity is imperfect,which greatly limits the molecular mechanism and the metabonomics development of A.niger YX-1217.The optimum genetic transformation of an industrial A.niger YX-1217 strain mediated by Agrobacterium tumefacien was determined with A.tumefaciens AGL-1 as the mediated strains,the initial A.niger YX-1217 spores concentration of 107 mL-1suspension,acetosyringone(AS)concentration of 300 μmol/L,and the volume ratio of the initial A.niger YX-1217 spores to A.tumefaciens of 1:9.Under such optimal transformation conditions,the transformation frequency was 25 transformants per plate using 107 conidia.Based on the genetic transformation,the application of A.niger YX-1217 as a high citric-acid production was expanded.The key gene cadA encoding conitate decarboxylase was overexpressed in the A.niger YX-1217 to increase itaconic acid production.Based on fed-batch fermentation,with three stage control of agitation speed,the highest itaconic acid production could reach 4.21 g/L with a productivity of 0.040 g/(L·h).Results showed the remarkable potential of A.niger YX-1217 to improve itaconic acid production.

参考文献/References:

[1] SOCCO C R,VANDENBERGHE L P S,RODRIGUES C,et al.New perspectives for citric acid production and application[J].Food technology biotechnology,2006,44(2):141.
[2] 满云,彭钢,陶惟一,等.色谱法提取柠檬酸的除杂工艺优化[J].生物加工过程,2017,15(3):29.
[3] MEYER V,MUELLER D,STROWIG T,et al.Comparison of different transformation methods for Aspergillus giganteus[J].Current genetics,2003,43(5):371.
[4] 郭艳梅,郑平,孙际宾.黑曲霉作为细胞工厂:知识准备与技术基础[J].生物工程学报,2010,26(10):1410.
[5] LI M.Establishment and optimization of Agrobacterium tumefaciensmediated transformation system of Aspergillus niger[J].China biotechnology,2012,32(1):56.
[6] 王艳,王文雅,袁其朋.根癌农杆菌介导又价载体pPK2转化黑曲霉条件的研究[J].食品科技,2013,38(8):56.
[7] XIE H,WANG F Q,WEI D Z.Morphological phenotypes of an Aspergillus niger strain with high citric acid production capacity[J].Research journal of biotechnology,2017,12(10):1.
[8] WILLKE T,VORLOP K D.Biotechnological production of itaconic acid[J].Applied microbiology and biotechnology,2001,56(3):289.
[9] LI A,VAN L N,TER B M,et al.A clone-based transcriptomics approach for the identification of genes relevant for itaconic acid production in Aspergillus[J].Fungal genetics biology,2011,48(6):602.
[10] LIN L,WANG F,WEI D.Chlorimuron ethyl as a new selectable marker for disrupting genes in the insect-pathogenic fungus Metarhizium robertsii[J].Journal of microbiology methods,2011,87(87):241.
[11] FANG W,YANG X,ZHANG Y,et al.Rapid extraction of DNA and RNA from fungi[J].Chinese journal of applied and environmental biology,2002,8:305.
[12] GANZLIN M,RINAS U.In-depth analysis of the Aspergillus niger glucoamylase(glaA)promoter performance using high-throughput screening and controlled bioreactor cultivation techniques[J].Journal of biotechnology,2008,135(3):266.
[13] KANAMASAS,DWIARTI L,OKABE M,et al.Cloning and functional characterization of the cis-aconitic acid decarboxylase(cad)gene from Aspergillus terreus[J].Applied microbiology and biotechnology,2008,80:223.
[14] MAO Y,YIN Y,ZHANG L,et al.Development of a novel Aspergillus,uracil deficient expression system and its application in expressing a cold-adapted α-amylase gene from Antarctic fungi Geomyces pannorum[J].Process biochemistry,2015,50(10),1581.
[15] OZEKI K,KYOYA F,HIZUME K,et al.Transformation of intact Aspergillus niger by electroporation[J].Bioscience biotechnology and biochemistry,1994,58(12):2224.
[16] 方卫国,张永军,杨星勇,等.根癌农杆菌介导真菌遗传转化的研究进展[J].中国生物工程杂志,2002,22(5):40.
[17] 亢新鑫,杨俊换,张怀渊,等.溶氧对耶氏解脂酵母油脂积累和柠檬酸分泌的影响[J].食品工业科技,2018,39(6):121.
[18] 李佳伟,刘龙,李江华,等.两阶段搅拌转速控制策略发酵生产柠檬酸[J].食品与生物技术学报,2014,33(2):131.
[19] HOSSAIN A H,LI A,BRICKWEDDE A,et al.Rewiring a secondary metabolite pathway towards itaconic acid production in Aspergillus niger[J].Microbial cell factories,2016,15(1):130.
[20] KUENZ A,GALLENMULLER Y,WILLKE T,et al.Microbial production of itaconic acid:developing a stable platform for high product concentrations[J].Applied microbiology and biotechnology,2012,96:1209.
[21] LI A,VANLUIJK N,BEEK-TER M,et al.A clonebased transcriptomics approach for the identification of genes relevant for itaconic acid production in Aspergillus[J].Fungal genetics biology,2011,48:602.
[22] YANG Z,GAO X,XIE H,et al.Enhanced itaconic acid production by self-assembly of two biosynthetic enzymes in Escherichia coli[J].Biotechnology bioengineering,2016,114(2):457.
[23] STEIGER M G,PUNT PJ,RAM A F J,et al.Characterizing MttA as a mitochondrial cis-aconitic acid transporter by metabolic engineering[J].Metabolic engineering,2016,35:95.

相似文献/References:

[1]李冬燕,于清跃.脱铝USY催化剂上柠檬酸三丁酯的合成[J].南京工业大学学报(自然科学版),2013,35(01):1.[doi:10.3969/j.issn.1671-7627.2013.01.020]
 LI Dongyan,YU Qingyue.Synthesis of tributyl citrate by dealuminium USY zeolite catalyst[J].Journal of NANJING TECH UNIVERSITY(NATURAL SCIENCE EDITION),2013,35(06):1.[doi:10.3969/j.issn.1671-7627.2013.01.020]
[2]满云,陶惟一,郭东升,等.柠檬酸发酵生产中玉米原料预处理工艺的条件优化[J].南京工业大学学报(自然科学版),2017,39(04):18.[doi:10.3969/j.issn.1671-7627.2017.04.004]
 MAN Yun,TAO Weiyi,LI Xiujuan,et al.Optimal conditions of corn pretreatment for citric acid fermentation[J].Journal of NANJING TECH UNIVERSITY(NATURAL SCIENCE EDITION),2017,39(06):18.[doi:10.3969/j.issn.1671-7627.2017.04.004]

备注/Memo

备注/Memo:
收稿日期:2018-05-21
基金项目:国家重大科技计划(2012ZX09304009)
作者简介:谢慧(1979—),女,河南郑州人,讲师,主要研究方向为工业微生物; 王风清(联系人),副教授,E-mail:fqwang@ecust.edu.cn.
引用本文:谢慧,马钦元,魏东芝,等.高产柠檬酸工业Aspergillus niger菌株转化体系构建及衣康酸发酵[J].南京工业大学学报(自然科学版),2018,40(6):40-50..
更新日期/Last Update: 2018-11-30