角蛋白降解菌Keratinibaculum paraultunense对废弃羽毛的发酵特性研究
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摘要
采用角蛋白降解菌Keratinibaculum paraultunense厌氧发酵废弃羽毛,以全培养基、无机盐培养基和纯水培养基分别进行发酵,测定了发酵过程中角蛋白酶的酶活、羽毛的降解率、可溶性蛋白和游离氨基酸的含量。研究发现,羽轴比羽枝更难被降解,但羽轴粉碎后,能达到同样的降解效果。采用无机盐培养基发酵时,主要降解产物是可溶性蛋白,含量可达0.93 mg/m L;而采用全培养基发酵时,主要发酵产物是角蛋白酶和游离氨基酸,当羽毛底物量提高到100 g/L,角蛋白酶的酶活高达11 179 U/m L。角蛋白降解菌K.paraultunense能快速高效地降解高浓度的废弃羽毛,可生产大量的可溶性蛋白和高活力的角蛋白酶。
Waste feathers were fermented at anaerobic condition using keratin degrading strain Keratinibaculum paraultunense in the whole medium,inorganic salt medium and pure water medium. The keratinase activity, feather degradation rate, the contents of soluble protein and free amino acids were determined during fermentation process. It was observed that pinna rachis was more difficult to be degraded than that of barbs. However, its degradation rate was comparable with that of barbs when pinna rachis was crushed. The main degradation product was soluble protein in the inorganic salt medium, and the concentration of soluble protein was 0.93 mg/ml. Meanwhile, the main degradation products were keratinase and free amino acids when using whole medium, and the activity of keratinase was 11 179 U/ml when feathers addition was increased to 100 g/L. The results indicated that K. paraultunense can be rapidly and effectively used to degrade high content of waste feathers, producing a large amount of soluble protein and keratinase with high activity.
Waste feathers were fermented at anaerobic condition using keratin degrading strain Keratinibaculum paraultunense in the whole medium,inorganic salt medium and pure water medium. The keratinase activity, feather degradation rate, the contents of soluble protein and free amino acids were determined during fermentation process. It was observed that pinna rachis was more difficult to be degraded than that of barbs. However, its degradation rate was comparable with that of barbs when pinna rachis was crushed. The main degradation product was soluble protein in the inorganic salt medium, and the concentration of soluble protein was 0.93 mg/ml. Meanwhile, the main degradation products were keratinase and free amino acids when using whole medium, and the activity of keratinase was 11 179 U/ml when feathers addition was increased to 100 g/L. The results indicated that K. paraultunense can be rapidly and effectively used to degrade high content of waste feathers, producing a large amount of soluble protein and keratinase with high activity.
引文
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[24]STIBOROVA H,BRANSKA B,VESELA T,et al.Transformation of raw feather waste into digestible peptides and amino acids[J].J Chem Technol Biot,2016,91(6):1629-1637.
[2]RAMAKRISHNA M R,SATHI K R,RANJITA Y C,et al.Effective feather degradation and keratinase production by Bacillus pumilus GRK for its application as bio-detergent additive[J].Bioresource Technol,2017,243(11):254-263.
[3]ABDELNABY M A,IBRAHIM M H,ELREFAI H A.Catalytic,kinetic and thermodynamic properties of Bacillus pumilus FH9 keratinase conjugated with activated pectin[J].Int J Biol Macromol,2016,85(9):238-245.
[4]蒋彪,王常高,杜馨,等.响应面法优化芽孢杆菌CJPE209产角蛋白酶发酵培养基的研究[J].中国酿造,2017,36(5):76-80.
[5]GOUSTEROVA A,BRAIKOVA D,GOSHEV I,et al.Degradation of keratin and collagen containing wastes by newly isolated thermoactinomycetes or by alkaline hydrolysis[J].Lett Appl Microbiol,2005,40(5):335-340.
[6]HE Z,SUN R,TANG Z,et al.Biodegradation of feather waste keratin by a keratin-degradingstrain of Bacillus subtilis[J].J Microb Biot,2018,28(2):314-322.
[7]SANGHVI G,PATEL H,VAISHNAV D,et al.A novel alkaline keratinase from Bacillus subtilis DP1 with potential utility in cosmetic formulation[J].Int J Biol Macromol,2016,87(3):256-262.
[8]RAJPUT R,GUPTA R.Thermostable keratinase from Bacillus pumilus KS12:production,chitin crosslinking and degradation of Sup35NM aggregates[J].Bioresource Technol,2013,133(4):118-126.
[9]KANCHANA R,MEATA D.Native feather degradation by a keratinophilic fungus[J].Int J Chemtech Res,2013,5(6):2947-2954.
[10]ELHOUL M B,JAOUADI N Z,REKIK H,et al.Biochemical and molecular characterization of new keratinoytic protease from Actinomadura viridilutea DZ50[J].Int J Biol Macromol,2016,92(7):299-315.
[11]ZAGHLOUL T I,EMBABY A M,ELMAHDY A R.Biodegradation of chicken feathers waste directed by Bacillus subtilis recombinant cells:scaling up in a laboratory scale fermentor[J].Bioresource Technol,2011,102(3):2387-2393.
[12]ZHANG R X,GONG J S,SU C,et al.Biochemical characterization of a novel surfactant-stable serine keratinase with no collagenase activity from Brevibacillus parabrevis CGMCC 10798[J].Int J Biol Macromol,2016,93(9):843-851.
[13]金扬,朱稳宇,韩瑞枝,等.角蛋白酶脱毛机理探究(一):脱毛条件的优化[J].皮革科学与工程,2011,21(3):14-17.
[14]PAUL T,DAS A,MANDAL A,et al.An efficient cloth cleaning properties of a crude keratinase combined with detergent:towards industrial viewpoint[J].J Clean Prod,2014,66(2):672-684.
[15]C魤LIN M,CONSTANTINESCU-ARUXANDEI D,ALEXANDRESCU E,et al.Degradation of keratin substrates by keratinolytic fungi[J].El J Biotechnol,2017,28(1):101-112.
[16]KITAGAWA S,MUKAI N,FURUKAWA Y,et al.Effect of soy peptide on brewing beer[J].J Biosci Bioeng,2008,105(4):360-366.
[17]HMIDET N,ALI N H,ZOUARI-FAKHFAKH N,et al.Chicken feathers:a complex substrate for the co-production of alpha-amylase and proteases by B.licheniformis NH1[J].J Ind Microbiol Biot,2010,37(9):983-990.
[18]SEDMAK J J,GROSSBERGS E.A rapid,sensitive and versatile assay for protein using Coomassie brilliant blue G250[J].Anal Biochem,2005,79(1-2):544-552.
[19]MAZOTTO A M,ASCHERI J L R,GODOY R L D O,et al.Production of feather protein hydrolyzed by B.subtilis,AMR and its application in a blend with cornmeal by extrusion[J].LWT-Food Sci Technol,2017,84(6):701-709.
[20]TESFAYE T,SITHOLE B,RAMJUGERNATH D,et al.Valorisation of chicken feathers:characterisation of physical properties and morphological structure[J].J Cl Prod,2017,149(4):349-365.
[21]GONG J,YUE W,ZHANG D,et al.A surfactant-stable Bacillus pumilus,K9-keratinase and its potential application in detergent industry[J].Chem Res Chinese U,2015,31(1):91-97.
[22]俞俊棠,唐孝宣,邬行彦,等.新编生物工艺学上册[M].北京:化学工业出版社,2003:165-166.
[23]MACIEL J L,WERLANG P O,DAROIT D J,et al.Characterization of protein-rich hydrolysates produced through microbial conversion of waste feathers[J].Waste Biomass Val,2017,8(4):1-10.
[24]STIBOROVA H,BRANSKA B,VESELA T,et al.Transformation of raw feather waste into digestible peptides and amino acids[J].J Chem Technol Biot,2016,91(6):1629-1637.