微生物學(xué)報(bào)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: [email protected]
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
假交替單胞菌Pseudoalteromonas sp. AJ5 產(chǎn)κ-卡拉膠酶的培養(yǎng)條件優(yōu)化
馬悅欣1, 2, 董雙林1, 劉雙連2, 牟海津3, 江曉路3*
(1 中國海洋大學(xué),，教育部海水養(yǎng)殖重點(diǎn)實(shí)驗(yàn)室, 青島266003)
(2 大連水產(chǎn)學(xué)院,，農(nóng)業(yè)部海洋水產(chǎn)增養(yǎng)殖學(xué)與生物技術(shù)重點(diǎn)開放實(shí)驗(yàn)室, 大連116023)
(3 中國海洋大學(xué)食品科學(xué)與工程學(xué)院, 青島266003)
摘要：【目的】本研究的目的是優(yōu)化Pseudoalteromonas sp. AJ5 菌株的培養(yǎng)條件使之產(chǎn)生高活性的胞外κ-卡拉膠酶?！痉椒ā客ㄟ^富集培養(yǎng)技術(shù)從刺參腸道分離出一株卡拉膠降解菌AJ5,，該菌株能利用卡拉膠作為惟一碳源和能源,。依據(jù)形態(tài)學(xué)和生理學(xué)特征及16S rRNA 基因序列分析，將該菌株鑒定為假交替單胞菌屬（ Pseudoalteromonas ）,。通過單因素試驗(yàn)和正交試驗(yàn)對Pseudoalteromonas sp. AJ5 菌株產(chǎn)胞外κ-卡拉膠酶的培養(yǎng)條件進(jìn)行了優(yōu)化,。【結(jié)果】單因素試驗(yàn)結(jié)果表明,，Pseudoalteromonas sp. AJ5 菌株的最佳培養(yǎng)條件為250 mL 三角瓶裝入75 mL 發(fā)酵培養(yǎng)基,、搖床轉(zhuǎn)速150 r/min、接種量7%,、pH8.0,。單因素試驗(yàn)和正交試驗(yàn)結(jié)果顯示該菌株的最佳培養(yǎng)基組成為κ-卡拉膠 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； 培養(yǎng)溫度為28℃,，培養(yǎng)時(shí)間為28 h?！窘Y(jié)論】Pseudoalteromonas sp. AJ5 菌株分泌胞外κ-卡拉膠酶,，在最佳培養(yǎng)條件下，該菌株的κ-卡拉膠酶活力比優(yōu)化前提高了4 倍,。
關(guān)鍵詞：κ-卡拉膠酶,；優(yōu)化；發(fā)酵條件,；培養(yǎng)基組成,；假交替單胞菌屬
中圖分類號： Q939.3 文獻(xiàn)標(biāo)識碼：A 文章編號：0001-6209(2008) 06-0764-08
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假交替單胞菌Pseudoalteromonas sp. AJ5 產(chǎn)κ-卡拉膠酶的培養(yǎng)條件優(yōu)化
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