微生物学报Acta Microbiologica Sinica 48(6)：757~764; 4 June 2008
Mariculture and Biotechnology Agriculture Ministry (K2006-12)
* Corresponding author. +86-532-82032290; Fax: +86-532-82032389; E-mail: jiangxl@ouc.edu.cn
Supported by the National Natural Science Foundation of China (40506027) and the Open Project of Program of Key Laboratory of Mariculture and Biotechnology Agriculture Ministry (K2006-12)
Optimization of κ-carrageenase production by Pseudoalteromonas sp. AJ5
Yuexin Ma1, 2, Shuanglin Dong1, Shuanglian Liu2, Haijin Mou3, Xiaolu Jiang3 *
(1 Key Laboratory of Mariculture, Ministry of Education, Ocean University of China, Qingdao 266003, China)
(2 Key Laboratory of Mariculture and Biotechnology Agriculture Ministry, Dalian Fisheries University, Dalian 116023, China)
(3 College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China)
Abstract: [Objectives] To optimize the culture conditions of Pseudoalteromonas sp. AJ5 for a higher production of extracellular κ-carrageenase. [Methods] A κ-carrageenan-degrading bacterium AJ5, capable of utilizing κ- carrageenan as sole source of carbon and energy, was isolated from the intestine of holothurian Apostichopus japonicus by enrichment culture technique. The strain was identified as the genus Pseudoalteromonas sp. according to its morphological and physiological characterization and 16S rRNA gene analysis. Culture conditions for the bacterium were standardized for the maximal productivity of the extracellular κ-carrageenase by the single factor and orthogonal tests. [Results] According to the single factor test, the optimal culture conditions were: 75 mL medium in 250 mL Erlenmeyer flask, shaking speed of 150 r/min, inoculum’s volume 7%, and pH 8.0. Based on the single factor and orthogonal tests the optimal medium components were: κ-carrageenan (1 g/L), beef extract (2 g/L ), NaCl (20 g/L), K2HPO4·3H2O (1 g/L), MgSO4·7H2O (0.5 g/L), MnCl2·4H2O (0.2 g/L), FePO4·4H2O (0.01 g/L), with the incubation temperature and time of 28°C and 28 h. [Conclusions] Pseudoalteromonas sp. AJ5 secreted an extracellular κ-carrageenase. Under the optimal culture conditions, four-fold increase in κ-carrageenase activity was achieved as compared to the control.
Keywords: κ-carrageenase; optimization; fermentation condition; medium component; Pseudoalteromonas
CLC number: Q939.3 Document code: A Article ID: 0001-6209(2008)06-0757-08
假交替单胞菌Pseudoalteromonas sp. AJ5 产κ-卡拉胶酶的培养条件优化
马悦欣1, 2, 董双林1, 刘双连2, 牟海津3, 江晓路3*
(1 中国海洋大学，教育部海水养殖重点实验室, 青岛266003)
(2 大连水产学院，农业部海洋水产增养殖学与生物技术重点开放实验室, 大连116023)
(3 中国海洋大学食品科学与工程学院, 青岛266003)
摘要：【目的】本研究的目的是优化Pseudoalteromonas sp. AJ5 菌株的培养条件使之产生高活性的胞外κ-卡拉胶酶。【方法】通过富集培养技术从刺参肠道分离出一株卡拉胶降解菌AJ5，该菌株能利用卡拉胶作为惟一碳源和能源。依据形态学和生理学特征及16S rRNA 基因序列分析，将该菌株鉴定为假交替单胞菌属（ Pseudoalteromonas ）。通过单因素试验和正交试验对Pseudoalteromonas sp. AJ5 菌株产胞外κ-卡拉胶酶的培养条件进行了优化。【结果】单因素试验结果表明，Pseudoalteromonas sp. AJ5 菌株的最佳培养条件为250 mL 三角瓶装入75 mL 发酵培养基、摇床转速150 r/min、接种量7%、pH8.0。单因素试验和正交试验结果显示该菌株的最佳培养基组成为κ-卡拉胶 1 g/L、牛肉膏2 g/L、 NaCl 20 g/L、K2HPO4·3H2O 1 g/L、 MgSO4·7H2O 0.5 g/L、MnCl2· 4H2O 0.2 g/L、 FePO4 · 4H2O 0.01 g/L； 培养温度为28℃，培养时间为28 h。【结论】Pseudoalteromonas sp. AJ5 菌株分泌胞外κ-卡拉胶酶，在最佳培养条件下，该菌株的κ-卡拉胶酶活力比优化前提高了4 倍。
关键词：κ-卡拉胶酶；优化；发酵条件；培养基组成；假交替单胞菌属
中图分类号： Q939.3 文献标识码：A 文章编号：0001-6209(2008) 06-0764-08
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