仿刺参消化道内产琼胶酶菌株的选育及培养条件优化Breeding and its optimal culture of bacterial and agarase-producing strain from digestive tracts in sea cucumber Apostichopus japonicus
马悦欣,安军,刘双连,徐高蓉,于书渤
摘要(Abstract)：
对仿刺参Apostichopus japonicus消化道内容物进行富集培养,以琼胶为惟一碳源进行平板初筛,摇瓶复筛,得到一株琼胶酶活性较高的菌株HF3。对该菌株进行紫外线诱变,获得突变菌株HF3-04,酶活力从43.0 U/mL增加到48.9 U/mL,提高了13.7%。通过正交试验,确定培养基的最佳组成为:琼胶3 g/L,硝酸铵1 g/L,磷酸氢二钠0.5 mmol/L,pH值8.0;最佳培养条件为:接种量为1%,250 mL三角瓶中发酵培养基的装液量为110 mL,摇床转速为150 r/m in,培养温度为25℃,培养时间为48 h。将培养基组成及培养条件优化后,菌株酶活力达到80.7 U/mL,比优化前提高了65.0%。
关键词(KeyWords)： 仿刺参;琼胶酶;紫外线诱变;培养条件优化
基金项目(Foundation): 辽宁省自然科学基金资助项目(20062129)
作者(Author): 马悦欣,安军,刘双连,徐高蓉,于书渤
DOI: 10.16535/j.cnki.dlhyxb.2007.02.002
参考文献(References)：
- [1]牟海津,江晓路,刘志鸿.利用微生物开发海洋多糖降解酶的研究[J].海洋科学,2003,27(11):10-13.
- [2]ARAKI C,ARAI K.Studies on the chemical constitution of agar-agar.XVⅢ.Isolation of a new crystalline disaccharide by enzy-matic hydrolysis of agar-agar[J].Bull Chem Soc Jpn,1956,29:339-345.
- [3]ARAKI T,ZHANG LU,MORISHITA T.Optimization of parame-ters for isolation of protoplasts fromGracilaria verrucosaRhodophy-ta[J].J Mar Biotechnol,1998(6):193-197.
- [4]汤海青,管斌,王雪鹏,等.琼胶降解菌AT-22的筛选和产酶性质[J].中国水产科学,2005,12(2):216-219.
- [5]杜宗军,王鹏,李筠.两株琼胶酶高产细菌的筛选和鉴定[J].海洋科学,2002,26(3):1-3.
- [6]褚艳,于文功,韩峰.琼胶酶高产海洋假单胞菌CY24的筛选及培养条件优化[J].中国海洋药物,2003(5):1-4.
- [7]AOKI T,ARAKI T,KITAMIKADO M.Purification and character-ization of a novel beta-agarase fromVibriosp.AP-2[J].Eur JBiochem,1990,187(2):461-465.
- [8]POTIN P,RICHARD C,ROCHAS C,et al.Purification and char-acterization of the alpha-agarase fromAlteromonas agarlyticus(Cataldi)comb.nov.,strain GJ1B[J].Eur J Biochern,1993,214(2):599-607.
- [9]VERA J,ALVAREZ R,MURANO E,et al.Identification of amarine agarolyticPseudoalteromonasand characterization of its ex-tracellular agarase[J].Appl Environ Microbiol,1998,64(11):4378-4383.
- [10]HA J C,KIMG T,KIMS K,et al.Beta-agarase fromPseudo-monassp.W7:purification of the recombinant enzyme fromEsche-richia coliand the effects of salt on its activity[J].Biotechnol ApplBiochem,1997,26:1-6.
- [11]ERASMUS J H,COOK P A,COYNE V E.The role of bacteriain the digestion of seaweed by the abaloneHaliotis midae[J].Aq-uaculture,1997,155:377-386.
- [12]中山大学生化教研室编.生化技术导论[M].北京:人民教育出版社,1978:53-55.
- [13]沈萍,范秀容,李广武.微生物学实验[M].北京:高等教育出版社,2000:124-127.
- [14]孙奕,陈騳.仿刺参体内外微生物组成及其生理特性的研究[J].海洋与湖沼,1989,20(4):300-307.
- [15]SAKAI T,ISHIZUKAK,KATO I.Isolation and characterizationof a fucoidan-degrading marine bacterium[J].Mar Biotechnol,2003,5(5):409-416.
- [16]陈惠源,蔡俊鹏,刘江涛.海洋细菌产琼胶酶的条件优化[J].现代食品科学技术,2005,21(3):48-50.
- [17]ΦSTGAARD K,WANGENB F,KNUTSENS H,et al.Large-scale production and purification ofκ-carrageenase fromPseudo-monas carageenovorafor applications in seaweed biotechnology[J].Enzyme Microb Technol,1993,15(4):326-333.