Constitutive expression of barley alpha-amylase in Pichia pastoris by high-density cell culture

文献类型： 外文期刊

第一作者： Liu, Z. W.

作者： Liu, Z. W.;Yin, H. X.;Yi, X. P.;Zhang, A. L.;Zhang, Z. H.;Shen, J. C.;Chen, L. P.;Luo, J. X.;Zhang, T. Y.;Luo, J. X.;Zhang, T. Y.;Fu, C. Y.

作者机构：

关键词： alpha-Amylase;Constitutive expression;Pichia pastoris;High-density cell culture;Bioactivity

期刊名称：MOLECULAR BIOLOGY REPORTS （ 影响因子：2.316； 五年影响因子：2.357 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： alpha-amy gene amplified from barley genome was cloned into MCS of pGAP9K to generate pGAP9K-alpha-amy which was then transformed into Pichia pastoris GS115 by electroporation. Transformants with multi-copies and high expression for the foreign gene were selected on G418 containing plate and expression analysis. The fermentation was carried out in a 50 l bioreactor with 20 l working volume, using a high-density cell culture method by continuously feeding with 50% glycerol-0.8% PTM4 to the growing culture for 54 h at 30A degrees C. Under the control of GAP promoter (pGAP), alpha-amy gene was constitutively expressed. At the end of the fermentation, the alpha-AMY expression reached 125 mg/l, while the biomass growth was 186 as measured by absorption of 600 nm. The secreted alpha-AMY was purified to 97.5% by SP-Sepharose FF ion-exchange chromatography and affinity purification. The recombinant alpha-AMY showed activity on hydrolysis of starch.

分类号： Q7

  • 相关文献

[1]Intracellular expression and purification of the Canstatin-N protein in Pichia pastoris. Yin, Huixiang,Liu, Zhenwang,Zhang, Ailian,Shen, Jincheng,Chen, Liping,Fu, Xian,Zhang, Zehua,Zhou, Bing,Fu, Ceyi,Zhang, Tianyuan,Luo, Jinxian,Zhang, Tianyuan,Luo, Jinxian.

[2]Inducible expression of human angiostatin by AOXI promoter in P. pastoris using high-density cell culture. Zhang, Ai-Lian,Qu, Zhi,Yi, Guo-Hui,Su, Dong-Xiao,Tu, Fa-Zhi,Pan, Ying-Wen,Zhang, Tian-Yuan,Luo, Jin-Xian,Zhang, Tian-Yuan,Luo, Jin-Xian,Fu, Ce-Yi.

[3]Constitutive expression of feedback-insensitive cystathionine gamma-synthase increases methionine levels in soybean leaves and seeds. Yu Yang,Hou Wen-sheng,Sun Shi,Wu Cun-xiang,Song Shi-kui,Han Tian-fu,Hacham, Yael,Matityahu, Ifat,Amir, Rachel,Hacham, Yael,Amir, Rachel. 2018

[4]Effect of 5 '-deletion of Arabidopsis profilin2 promoter on its vascular specific expression. Liu, YH,Wang, ZX,Jia, SR. 2001

[5]Noninvasive Expressions of ipt in Whole Plants or Roots through pOp/LhG4 Indicate a Role of Plant Aerial Parts and Light in Cytokinin Synthesis and Root Inhibition. Guo, Jianchun,Hu, Xinwen.

[6]Ability of resveratrol to inhibit advanced glycation end product formation and carbohydrate-hydrolyzing enzyme activity, and to conjugate methylglyoxal. Shen, Yixiao,Sheng, Zhanwu,Shen, Yixiao,Xu, Zhimin. 2017

[7]Screening, Gene Cloning, and Characterizations of an Acid-Stable alpha-Amylase. Liu, Xinyu,An, Yi,Cheng, Kun,Wang, Mingdao,Yang, Sen,Chen, Hongge,Jia, Wei.

[8]Isolation of -Amylase Inhibitors from Kadsura longipedunculata Using a High-Speed Counter-Current Chromatography Target Guided by Centrifugal Ultrafiltration with LC-MS. Cen, Yin,Chen, Xiaoqing,Xiao, Aiping,Liu, Liangliang.

[9]Improving the Catalytic Performance of a Talaromyces leycettanus alpha-Amylase by Changing the Linker Length. Zhang, Duoduo,Luo, Xuegang,Zhang, Tongcun,Zhang, Duoduo,Luo, Xuegang,Zhang, Tongcun,Tu, Tao,Wang, Yuan,Li, Yeqing,Zheng, Fei,Wang, Xiaoyu,Bai, Yingguo,Huang, Huoqing,Su, Xiaoyun,Yao, Bin,Luo, Huiying.

[10]Seed micromorphology and germination characteristics of wild and cultivated pepper strains. Zhang Zhuqing,Dai Xiongze,Zou Xuexiao,Ou Lijun,Ou Lijun.

[11]Phytosterols in banana (Musa spp.) flower inhibit alpha-glucosidase and alpha-amylase hydrolysations and glycation reaction. Sheng, Zhanwu,Pan, Siyi,Sheng, Zhanwu,Ai, Binling,Zheng, Lili,Zheng, Xiaoyan,Dai, Haofu,Prinyawiwatkul, Witoon,Xu, Zhimin.

[12]Screening and Binding Analysis of Flavonoids with Alpha-Amylase Inhibitory Activity from Lotus Leaf. Liao, Liping,Chen, Jing,Liu, Liangliang,Xiao, Aiping. 2018

[13]Physicochemical characterization of enzymatically hydrolyzed heat treated granular starches. Mu, Tai-Hua,Abegunde, Oluwaseyi K.,Sun, Hong-Nan,Deng, Fu-Ming,Zhang, Miao.

[14]Overexpression of celB gene coding for beta-glucosidase from Pyrococcus furiosus using a baculovirus expression vector system in silkworm, Bombyx mori. Lin, Xu'Ai,Zhang, Wei,Chen, Yin,Bin Yao,Zhang, Zhi Fang.

[15]Analysis of alpha-amylase inhibitor from corni fructus by coupling magnetic cross-linked enzyme aggregates of alpha-amylase with HPLC-MS. Cen, Yin,Liu, Fang,Yu, Jingang,Jiang, Xinyu,Chen, Xiaoqing,Liu, Liangliang.

[16]Screening and separation of alpha-amylase inhibitors from Solanum nigrum with amylase-functionalized magnetic graphene oxide combined with high-speed counter-current chromatography. Cen, Yin,Chen, Xiaoqing,Xiao, Aiping,Liu, Liangliang. 2017

[17]Study on the interaction between 3 flavonoid compounds and alpha-amylase by fluorescence spectroscopy and enzymatic kinetics. Li, Y.,Gao, F.,Gao, F.,Zhao, C.,Shan, F.,Bian, J..

[18]Germplasm improvement for preharvest sprouting resistance in Chinese white-grained wheat: An overview of the current strategy. Xiao, SH,Zhang, XY,Yan, CS,Lin, H. 2002

[19]Preparation and Characterization of Nano Fe3O4/Carboxylation Chitosan/Alginate Sodium Magnetofluid by One Pot Method. Jiang, Jianfang,He, Chunyang,Xiong, Jun,Tu, Wenya,Jiang, Hao. 2016

[20]Effect of salinity on the rearing performance of juvenile golden pompano Trachinotus ovatus (Linnaeus 1758). Ma, Zhenhua,Guo, Huayang,Zheng, Panlong,Wang, Long,Jiang, Shigui,Zhang, Dianchang,Ma, Zhenhua,Ma, Zhenhua,Qin, Jian G..

作者其他论文 更多>>

微信小程序