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高渗条件下利用蔗糖提升2-酮基-L-古龙酸生产效率
引用本文:陈克杰,周景文,刘立明,刘杰,堵国成,陈坚. 高渗条件下利用蔗糖提升2-酮基-L-古龙酸生产效率[J]. 生物工程学报, 2010, 26(11): 1507-1513
作者姓名:陈克杰  周景文  刘立明  刘杰  堵国成  陈坚
作者单位:1. 江南大学食品科学与技术国家重点实验室,无锡214122;江南大学工业生物技术教育部重点实验室,无锡214122
2. 江苏江山制药有限公司,靖江,214500
3. 江南大学工业生物技术教育部重点实验室,无锡,214122
基金项目:国家高技术研究发展计划 (863计划) (No. 2006AA020303),“十一五”国家科技支撑计划 (No. 2007BAI46B02),国家重点基础研究发展计划(973计划) (No. 2007CB714306),国家自然科学基金重点项目 (No. 20836003) 资助。
摘    要:旨在进一步提升维生素C前体2-酮基-L-古龙酸(2-KLG)的生产效率。在详细考察了2-KLG工业化生产过程中渗透压变化规律的基础上,研究了高渗对混合菌系细胞生长和2-KLG合成的影响,提出蔗糖促进伴生菌巨大芽胞杆菌Bacillus megaterium生长,进而促进普通生酮古龙酸菌Ketogulonigenium vulgare生长和产酸的策略。结果表明,2-KLG的积累和碱性物质的流加使渗透压上升了832mOsmol/kg;高渗抑制了巨大芽胞杆菌的生长(15.4%),从而抑制普通生酮古龙酸菌(31.7%)的生长,导致2-KLG产量和生产强度分别下降67.5%和69.3%(以1250mOsmol/kg为例);蔗糖的添加则显著促进巨大芽胞杆菌的生长,使高渗条件下(摇瓶,1250 mOsmol/kg)2-KLG产量(40.6g/L)提高87%;在3L发酵罐中,补加10mmol/L蔗糖使2-KLG发酵周期缩短10.8%,2-KLG生产强度提高10.4%。研究成果为在环境胁迫下提高混菌生产目标代谢产物的产量提供了潜在的策略。

关 键 词:混菌发酵,2-酮基-L-古龙酸,碳源调节,维生素C,高渗
收稿时间:2010-04-16

Enhancing 2-keto-L-gulonic acid production under hyperosmotic stress by adding sucrose
Kejie Chen,Jingwen Zhou,Liming Liu,Jie Liu,Guocheng Du and Jian Chen. Enhancing 2-keto-L-gulonic acid production under hyperosmotic stress by adding sucrose[J]. Chinese journal of biotechnology, 2010, 26(11): 1507-1513
Authors:Kejie Chen  Jingwen Zhou  Liming Liu  Jie Liu  Guocheng Du  Jian Chen
Affiliation:State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China; Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China; Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China; Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;Jiangsu Jiangshan Pharmaceutical Co. Ltd., Jingjiang 214500, China;Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China;State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China; Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China
Abstract:This study aimed to further enhance 2-keto-L-gulonic acid (2-KLG) production efficiency. A strategy for enhancing Ketogulonigenium vulgare growth and 2-KLG production by improving B. megaterium growth with sucrose was developed based on the time course of osmolality during 2-KLG industrial scale fermentation and effects of osmolality on cells growth and 2-KLG production. Results showed that the accumulation of 2-KLG and the feeding of alkaline matter led to an osmolality rise of 832 mOsmol/kg in the culture broth. High osmotic stress (1 250 mOsmol/kg) made the growth of B. megaterium and K. vulgare decreased 15.4% and 31.7%, respectively, and consequently the titer and productivity of 2-KLG reduced 67.5% and 69.3%, respectively. When supplement sucrose under high osmotic condition (1 250 mOsmol/kg), B. megaterium growth was significantly improved, with the result that 2-KLG production was increased 87%. Furthermore, by applying this sucrose addition strategy further to batch fermentation in 3 L fermentor, the productivity of 2-KLG increased 10.4%, and the duration of fermentation declined 10.8%. The results presented here provide a potential strategy for enhancing the target metabolites produced by mixed strains at environmental stress.
Keywords:mixed culture   2-keto-L-gulonic acid   carbon source regulation   Vitamin C   hyperosmotic stress
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