为筛选1株产蛋白酶的海洋菌种,课题组从厦门市红树林中筛选得到1株产蛋白酶的丝状真菌,并对其细胞形态进行了扫描电镜观察,经提取其ITS序列于Genbank进行比对并构建进化树,将其鉴定为Parengyodontium album HX2019006。该菌株在海水培养基中可生产蛋白酶,通过对其培养基组分及发酵条件进行单因素优化,获得较高的蛋白酶产量后,选择培养基中麦芽糖浓度、蛋白胨浓度和培养时间3个因素进行3水平响应面优化,在优化后的培养基和发酵条件下,发酵液蛋白酶活力为71.1 U/mL,较初始条件提高125.7%。
傅奇
,
林俊杰
,
庄峙厦
,
黄华斌
,
孙恩贤
,
甘美裕
,
肖玉娟
. 一株海洋来源蛋白酶产生菌Parengyodontium album HX2019006的鉴定及其发酵条件研究[J]. 食品与发酵工业, 2020
, 46(10)
: 185
-190
.
DOI: 10.13995/j.cnki.11-1802/ts.023581
A marine protease-producing fungal strain was screened from the local sea sediments in Xiamen. The morphology of this fungal strain was observed by scanning electron microscopy, and it was identified as Parengyodontium album HX2019006 by extracting the ITS sequence and comparing it in Genbank with the subsequent generation of the phylogenetic tree. This strain was capable of producing protease in seawater-based culture. We performed a single-factor optimization of the culture medium components and fermentation conditions to achieve an increase in protease yield. Three levels of maltose concentration, peptone concentration, and culture time were selected for the response surface methodology (RSM) optimization. Under the optimized culture medium and fermentation conditions, the protease activity in the fermentation broth was 71.1 U/mL, which was 125.7% higher than that under the initial conditions.
[1] CHI Z, MA C, WANG P, et al. Optimization of medium and cultivation conditions for alkaline protease production by the marine yeast Aureobasidium pullulans[J]. Bioresource Technology, 2007, 98(3): 534-538.
[2] ZHANG S C, SUN M, LI T, et al. Structure analysis of a new psychrophilic marine protease[J]. PLoS One, 2011, 6(11):26 939.
[3] RAMESH S, RAJESH M, MATHIVANAN N. Characterization of a thermostable alkaline protease produced by marine Streptomyces fungicidicus MML1614[J]. Bioprocess and biosystems engineering, 2009, 32(6): 791-800.
[4] KUMAR C G, JOO H S, KOO Y M, et al. Thermostable alkaline protease from a novel marine haloalkalophilic Bacillus clausii isolate[J]. World Journal of Microbiology and Biotechnology, 2004, 20(4): 351-357.
[5] SOMERFIELD P J, CLARKE K R. A comparison of some methods commonly used for the collection of sublittoral sediments and their associated fauna[J]. Marine Environmental Research, 1997, 43(3): 145-156.
[6] 李震, 肖秧, 甄天元, 等. 海参肠道产蛋白酶菌的分离鉴定及其在海参下脚料中的应用[J]. 食品与发酵工业, 2019, 45(18): 195-201.
[7] 侯靖, 许文梅, 崔恒林. 柴达敏盐湖产胞外蛋白酶嗜盐古菌的筛选及其酶学性质[J]. 食品与发酵工业, 2018, 44(3): 91-96.
[8] HALL T A. Bioedit: A user—friendly biological sequence alignment editor and analysis program for Windows 95/98/NT[J]. Nucleic Acids Symposium Series, 1999, 41:95-98.
[9] THOMSON J D, GIBSON T J, PLEWNIAK F, et al. The Clustal_X windows interface: Flexible strategies for multiple sequence alignment aided by quality analysis tools[J]. Nucleic Acids Research. 1997,25:4 876-4 882.
[10] JIANG X Z, YU H Y, XIANG M C, et al. Echinochlamydosporium variabile, a new genus and species of Zygomycota from soil nematodes[J]. Fungal Divers, 2011,46(1):43-51.
[11] 肖玉娟, 傅奇, 沈金海, 等. 一株引起厦门地区不同作物感染的 Poitrasia circinans分离与鉴定[J]. 科学技术与工程, 2018, 18(14): 20-23.
[12] CHAVERRI P, SAMUELS G J, STEWART E L, et al. Hypocrea nigrovirens, a new species with a gliocladium-like anamorph[J]. Mycologia, 2001, 93(4): 758-763.
[13] GB/T 23527—2009蛋白酶制剂[S]. 北京: 中国标准出版社, 2009.
[14] 张若兰, 刘庆国, 王敏, 等. 枯草芽孢杆菌碱性蛋白酶基因在酿酒酵母中的表达和应用[J]. 食品与发酵工业, 2018, 44(7): 76-81.
[15] 蔺志颖, 张博文, 李佳. 响应面法优化椰浆蛋白发酵饮料发酵工艺[J]. 中国酿造, 2019,38(5): 210-214.
[16] KORTING H C, SCHALLER M. Neue Entwicklungen in der Mykologie[J]. Hautarzt,2001,52(2):91-97.
[17] NAM K H. Triglycine-based approach for identifying the substrate recognition site of an enzyme[J]. Crystals, 2019, 9(9): 444.
[18] REN Y, LUO H, HUANG H, et al. Improving the catalytic performance of Proteinase K from Parengyodontium album for use in feather degradation[J]. International Journal of Biological Macromolecules, 2019,DOI:10.1016/j.ijbiomac.2019.11.043.
[19] SAMAL B B, KARAN B, BOONE T C, et al. Isolation and characterization of the gene encoding a novel, thermostable serine proteinase from the mould Tritirachium album Limber[J]. Molecular microbiology, 1990, 4(10): 1 789-1 792.
[20] CHELLAPPAN S, JASMIN C, BASHEER S M, et al. Characterization of an extracellular alkaline serine protease from marine BTMFS10[J]. Journal of industrial microbiology & biotechnology, 2011, 38(6): 743-752.
[21] 沈萍, 陈向东. 微生物学[M]. 北京: 高等教育出版社, 2016: 84-138.
[22] GOLDMAN J C, CARON D A, DENNETT M R. Regulation of gross growth efficiency and ammonium regeneration in bacteria by substrate C: N ratio 1[J]. Limnology and Oceanography, 1987, 32(6): 1 239-1 252.
[23] EBELING W, HENNRICH N, KLOCKOW M, et al. Proteinase K from Tritirachium album limber[J]. European Journal of Biochemistry, 1974, 47(1): 91-97.
[24] FORTELIUS C, MARKKANEN P. Nutritional regulation of proteinase production in the fungus, Tritirachium album[J]. Journal of Industrial Microbiology and Biotechnology, 2000, 24(6): 369-373.
[25] TSANG C C, CHAN J F W, PONG W M, et al. Cutaneous hyalohyphomycosis due to Parengyodontium album gen. et comb. nov[J]. Medical Mycology, 2016, 54(7): 699-713.
[26] 陈坚, 堵国成. 发酵工程原理与技术[M]. 北京: 化学工业出版社, 2018.
