Enhanced Riboflavin Production by Expressing Heterologous Riboflavin Operon from B.cereus ATCC14579 in Bacillus subtilis

CIESC Journal ›› 2010, Vol. 18 ›› Issue (1): 129-136.

Enhanced Riboflavin Production by Expressing Heterologous Riboflavin Operon from B.cereus ATCC14579 in Bacillus subtilis

段云霞^1,2,3, 陈涛^1,2, 陈洵^1,2, 王靖宇^1,2, 赵学明^1,2

1. Department of Biochemical Engineering, School of Chemical Engineering and Technology, Edinburgh-Tianjin Joint Research Center for System Biology and Synthetic Biology, Tianjin University, Tianjin 300072, China;
2. Key Laboratory of Systems Bioengineering, Ministry of Education, Tianjin 300072, China;
3. Tianjin Academy of Environmental Sciences, Tianjin 300196, China

收稿日期:2009-03-20 修回日期:2009-07-22 出版日期:2010-02-28 发布日期:2010-12-30
通讯作者: CHEN Tao, E-mail: chentao@tju.edu.cn
作者简介:
基金资助:
Supported by the National Natural Science Foundation of China(20536040);the State Key Development Program for Basic Research of China(2007CB707802);the Development Project of Science and Technology of Tianjin(05YFGZGX04500)

Enhanced Riboflavin Production by Expressing Heterologous Riboflavin Operon from B.cereus ATCC14579 in Bacillus subtilis

DUAN Yunxia^1,2,3, CHEN Tao^1,2, CHEN Xun^1,2, Wang Jingyu^1,2, ZHAO Xueming^1,2

1. Department of Biochemical Engineering, School of Chemical Engineering and Technology, Edinburgh-Tianjin Joint Research Center for System Biology and Synthetic Biology, Tianjin University, Tianjin 300072, China;
2. Key Laboratory of Systems Bioengineering, Ministry of Education, Tianjin 300072, China;
3. Tianjin Academy of Environmental Sciences, Tianjin 300196, China

Received:2009-03-20 Revised:2009-07-22 Online:2010-02-28 Published:2010-12-30
Supported by:
Supported by the National Natural Science Foundation of China(20536040);the State Key Development Program for Basic Research of China(2007CB707802);the Development Project of Science and Technology of Tianjin(05YFGZGX04500)

摘要/Abstract

摘要： Fragment containing the whole riboflavin(rib)operons of B.cereus ATCC14579 was detected from GenBank and annotated.The rib operon of ATCC14579 was cloned with Pn,its native promoter,or with P43,the vegetative growth promoter,into the plasmid.Expression analysis showed that heterologous rib operon was operative in B.subtilis.Integrative plasmid with P43-rib fragment was integrated into the chromosome of B.subtilis RH33,yielding transformant B.subtilis PY.With optimized medium components,4.3 g·L^-1 of riboflavin was achieved in batch culture of B.subtilis PY,which was 27%enhancement compared to the host strain.Real-time reverse transcription polymerase chain reaction(RT-PCR)analysis indicated that the transcriptional level of ribA maintained 2.8-fold higher with the expression of herterologous riboperon.Furthermore,the stability of B.subtilis PY was increased form 45%to 87%.The high transcriptional level of rib gene and higher stability of B.subtilis PY could explain the increased riboflavin production.

关键词: iosynthesis of riboflavin, heterologous gene expression, P43 promoter, riboflavin(rib)operon, Bacillus subtilis

Abstract: Fragment containing the whole riboflavin(rib)operons of B.cereus ATCC14579 was detected from GenBank and annotated.The rib operon of ATCC14579 was cloned with Pn,its native promoter,or with P43,the vegetative growth promoter,into the plasmid.Expression analysis showed that heterologous rib operon was operative in B.subtilis.Integrative plasmid with P43-rib fragment was integrated into the chromosome of B.subtilis RH33,yielding transformant B.subtilis PY.With optimized medium components,4.3 g·L^-1 of riboflavin was achieved in batch culture of B.subtilis PY,which was 27%enhancement compared to the host strain.Real-time reverse transcription polymerase chain reaction(RT-PCR)analysis indicated that the transcriptional level of ribA maintained 2.8-fold higher with the expression of herterologous riboperon.Furthermore,the stability of B.subtilis PY was increased form 45%to 87%.The high transcriptional level of rib gene and higher stability of B.subtilis PY could explain the increased riboflavin production.

Key words: iosynthesis of riboflavin, heterologous gene expression, P43 promoter, riboflavin(rib)operon, Bacillus subtilis

段云霞, 陈涛, 陈洵, 王靖宇, 赵学明. Enhanced Riboflavin Production by Expressing Heterologous Riboflavin Operon from B.cereus ATCC14579 in Bacillus subtilis[J]. CIESC Journal, 2010, 18(1): 129-136.

DUAN Yunxia, CHEN Tao, CHEN Xun, Wang Jingyu, ZHAO Xueming. Enhanced Riboflavin Production by Expressing Heterologous Riboflavin Operon from B.cereus ATCC14579 in Bacillus subtilis[J]. , 2010, 18(1): 129-136.

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Enhanced Riboflavin Production by Expressing Heterologous Riboflavin Operon from B.cereus ATCC14579 in Bacillus subtilis

Enhanced Riboflavin Production by Expressing Heterologous Riboflavin Operon from B.cereus ATCC14579 in Bacillus subtilis

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