[1]裘梁,杨萍,董德刚,等.重组烟曲霉壳聚糖酶在毕赤酵母中高密度发酵表达及性质研究[J].江西师范大学学报(自然科学版),2013,(06):607-610.
 QIU Liang,YANG Ping,DONG De-gang,et al.The Study on High Density Fermentation of Recombinant Chitosanase from Aspergillus fumigatus in Pichia pastoris and Characterization of Recombinant Chitosanase[J].Journal of Jiangxi Normal University:Natural Science Edition,2013,(06):607-610.
点击复制

重组烟曲霉壳聚糖酶在毕赤酵母中高密度发酵表达及性质研究()
分享到:

《江西师范大学学报》(自然科学版)[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2013年06期
页码:
607-610
栏目:
出版日期:
2013-12-31

文章信息/Info

Title:
The Study on High Density Fermentation of Recombinant Chitosanase from Aspergillus fumigatus in Pichia pastoris and Characterization of Recombinant Chitosanase
作者:
裘梁;杨萍;董德刚;阳历;王曼莹
江西中医药大学生物技术研发中心,江西南昌,330096;江西师范大学生命科学学院,江西省食品与生物技术产学研合作示范基地,江西南昌330022;江西中医药大学生物技术研发中心,江西南昌330096;江西师范大学生命科学学院,江西省食品与生物技术产学研合作示范基地,江西南昌330022
Author(s):
QIU Liang;YANG Ping;DONG De-gang;YANG Li;WANG Man-ying
关键词:
重组壳聚糖酶毕赤酵母高密度发酵
Keywords:
recombinant chitosanasePichia pastorishigh density fermentation
分类号:
Q786
文献标志码:
A
摘要:
采用高密度发酵法在5L发酵罐中对pPIC9-csn/GS115重组毕赤酵母Pichia pastoris菌株进行发酵,壳聚糖酶的产量达到4200 U·mL-1,发酵液经乙醇浓缩、透析和阳离子交换色谱等步骤进行分离纯化,获得了纯度较高的重组壳聚糖酶.该重组壳聚糖酶与野生型壳聚糖酶对壳聚糖作用模式相同,其最终产物均为壳三糖,且其最适作用温度为60℃,最适pH值为6.0.
Abstract:
The recombinant Pichia pastoria strain infected with pPIC9-csn/GS115 for expression of chitosanase was high cell density cultured in a 5 L fermentor.As a result,the yield of chitosanase was 4 200 U·mL,the chitosanase fermentation product was purified to above 95% by ethanol concentration,dialysis and cation exchange chromatography.In addition,the recombinant chitosanase showed the same mode of action as the wild chitosanase,produced the final product chitotriose and were optimal at the temperature of 60 ℃ and pH of 6.0.

参考文献/References:

[1] Yoshihara K,Hosokawa J,Kubo T,et al.Purification and properties of a chitosanase from Pseudomonas sp.H-14 [J].Biosci Biotech Biochem,1992,56(6):972-973.
[2] 王超莉,李扬忠.羟乙基壳聚糖硫酸酯的合成与表征 [J].江西师范大学学报:自然科学版,2009,33(3):272-274.
[3] Yamasaki Y,Hayashi I,Ohta Y,et al.Purification and mode of action of chitosanolytic enzyme from Enterobacter sp.G-1 [J].Biosci Biotechnol Biochem,1993,57(3):444-449.
[4] Omumasaba C A,Yoshida N,Sekiguchi Y,et al.Purification and some properties of a novel chitosanase from Bacillus subtilis KH1 [J].Appl Microbiol,1999,46(1):162-271.
[5] Shimosaka M,Nogawa M,Ohino Y.Chitosanase from the plant pathogenic fungus Fusarium solanif sp.phaseoli-purification and some properties [J].Biosci Biotechnol Biochem,1993,57(2):231-235.
[6] Somasherkar D,Joseph R.Chitosanases:properties and applications:a review [J].Bioresource Technology,1996,55(1):35-45.
[7] Liu Jing,Xia Wenshui.Purification and characterization of a bifunctional enzyme with chitosanase and cellulase activity from Commercial cellulose [J].Biochemical Engineering Journal,2006,30(1):82-87.
[8] Chen Yaling,Su Chiakai,Chiang Beenhuang.Optimization of reversed micellar extraction of chitosanases produced by Bacillus cereus [J].Process Biochemistry,2006,41(4):752-758.
[9] Chen Xiaoe,Xia Wenshui,Yu Xiaobin.Purification and characterization of two types of chitosanase from Aspergillus sp.[J].Food Research International,2005,38(3):315-322.
[10] Kim P I,Kang T H,Chung K J,et al.Purification of a constitutive chitosanase produced by Bacillus sp.MET 1299 with cloning and expression of the gene FEMS [J].Microbiology Letters,2004,240(1):31-39.
[11] Johnsen M G,Hansen O C,Stougaard P.Isolation,characterization and heterologous expression of a novel chitosanase from Janthinobacterium sp.strain 4239 [J].Microbial Cell Factories,2010,9(1):5.
[12] 梁东春,左爱军,郭刚,等.烟曲霉菌壳聚糖酶基因的克隆及在大肠杆菌中的表达 [J].微生物学报,2005,45(4):539-542.
[13] Zhang Jiquan,Sun Yuying.Molecular cloning,expression and characterization of a chitosanase from Microbacterium sp.[J].Biotechnol Lett,2007,29(8):1221-1225.
[14] 杨萍.烟曲霉壳聚糖酶基因的克隆与毕赤酵母表达 [D].南昌:江西师范大学,2007.
[15] 裘梁,杨萍,游清徽,等.密码子优化烟曲霉壳聚糖内切酶基因在毕赤酵母中的高效表达 [J].安徽农业科学,2012,40(26):12769-12771.
[16] Gregg J M,Vedvick T S,Raschke W C.Recent advances in the expression of foreign genes Pichia pastoris [J].Biotechnology,1993,11(8):905-910.

备注/Memo

备注/Memo:
国家自然科学基金(81260604);江西省教育厅青年科学基金(GJJ11195)
更新日期/Last Update: 1900-01-01