ارزیابی تولید آنزیم ترنس گلوتامیناز در جدایه استرپتومیست بومی جدا شده از خاک‌های کشاورزی استان قم
محورهای موضوعی :
میکروب شناسی
سمیه بحری
1
,
سید سهیل آقائی
2
,
سید محمدحسین رضویان
3
,
فائزه کبیری
4
1 - کارشناسی ارشد، گروه زیست‌شناسی، دانشکده علوم پایه، واحد قم، دانشگاه آزاد اسلامی، قم، ایران.
2 - استادیار، گروه میکروبیولوژی، دانشکده علوم پایه، واحد قم، دانشگاه آزاد اسلامی، قم، ایران
3 - دانشیار، گروه میکروبیولوژی، دانشکده علوم پایه، واحد قم، دانشگاه آزاد اسلامی، قم، ایران.
4 - دکتری، گروه میکروبیولوژی، دانشکده علوم پایه، واحد قم، دانشگاه آزاد اسلامی، قم، ایران
تاریخ دریافت : 1402/02/07
تاریخ پذیرش : 1402/04/13
تاریخ انتشار : 1401/10/01
کلید واژه:
آنزیم,
ریزوسفر,
استرپتومیست,
آکتینوباکتری,
ترنس گلوتامیناز,
چکیده مقاله :
هدف: آنزیم ترنس گلوتامیناز نوعی آنزیم پروتئاز از نوع آسیل ترانسفراز است که پیوندهای تشکیل شده توسط این آنزیم سبب ایجاد استحکام و پایداری در محصولات صنایع مختلف می‌گردد. آکتینوباکتری‌ها به دلیل توانایی در تولید متابولیت‌های ثانویه و آنزیم‌ها، دارای کاربرد زیست فناورانه هستند. مطالعه حاضر با هدف شناسایی و غربالگری سویه‌های آکتینوباکتری مولد ترنس گلوتامیناز، از خاک‌های ناحیه ریزوسفری گیاهان دارویی بومی استان قم انجام شد.مواد و روش‌ها: در این تحقیق 100 نمونه از خاک مزارع کشاورزی استان قم در شرایط کاملاً استریل جمع‌آوری گردید و پس از تهیه رقت‌های متوالی در سطح محیط کشت ISP3 آگار، کشت داده شد. جدایه‌های آکتینوباکتری توسط شناساگر اختصاصی آنزیم ترنس گلوتامیناز غربالگری شدند. فاکتورهای محیطی شامل محیط کشت و دور شیکر بر رشد جدایه و تولید آنزیم، بهینه‌سازی گردید. به منظور شناسایی و بررسی فیلوژنی جدایه آکتینوباکتری، از روش‌های فنوتیپی و مولکولی استفاده شد.یافته‌ها: از میان 40 جدایه آکتینوباکتری، 5 سویه، براساس هاله تغییر رنگ توسط سوبسترای اختصاصی N-Carbobenzoxy (CBZ)، توانایی تولید آنزیم را نشان دادند. جدایه منتخب با بیشترین میزان تولید آنزیم، از نظر ترادف ژن 16S rRNA، با جنس استرپتومایسس شباهت داشت.نتیجه‌گیری: نتایج این مطالعه نشان داد که جدایه استرپتومیست بومی خاک‌های کشاورزی استان قم، دارای توانایی قابل ملاحظه‌ای برای تولید آنزیم ترنس گلوتامیناز است که برای اولین بار مورد مطالعه قرار گرفت. این جدایه می‌تواند در صنایع غذایی و دارویی مختلف جهت تولید آنزیم ترنس گلوتامیناز استفادهشود.
چکیده انگلیسی:
Purpose: Transglutaminase enzyme is a type of protease enzyme of acyltransferase type, the bonds formed by this enzyme cause strength and stability in the products of various industries. Actinobacteria have biotechnological applications due to their ability to produce secondary metabolites and enzymes. The present study was conducted with the aim of identifying and screening trans-glutaminase-producing actinobacterial strains from the soils of the rhizosphere area of native medicinal plants in Qom province.Materials and Methods: In this research, 100 samples of soil from agricultural fields in Qom province were collected in completely sterile conditions and after preparing successive dilutions, they were cultured on ISP3 agar culture medium. Actinobacterial isolates were screened by transglutaminase enzyme specific identifier. Environmental factors including culture medium and shaker cycle were optimized on isolate growth and enzyme production. In order to identify and investigate the phylogeny of actinobacterial isolates, phenotypic and molecular methods were used.Findings: Out of 40 actinobacterial isolates, 5 strains showed the ability to produce the enzyme, based on the halo of color change by the specific substrate N-Carbobenzoxy (CBZ). The selected isolate with the highest amount of enzyme production, in terms of 16S rRNA gene synonymy, was similar to Streptomyces genus.Conclusion: The results of this study showed that Streptomyces isolate native to agricultural soils of Qom province has a significant ability to produce transglutaminase enzyme, which was studied for the first time. This isolate can be used in various food and pharmaceutical industries to produce transglutaminase enzyme.
منابع و مأخذ:
Abd-Rabo F, Ei-Dieb S, Abd-EI-Fattah A & Sakr S. Natural state changes of cows' and buffaloes' milk proteins induced by microbial transglutaminase. J Am Sci. 2010; 6(9): 612-20.
Bourneow C, Benjakul S & H-Kittikun A. Hydroxamate-based colorimetric method for direct screening of transglutaminase-producing bacteria. World Journal of Microbiology and Biotechnology. 2012; 28: 2273-7.
Bahrim G, Iancu C, Buţu N & Negoiţă TG. Production of a novel microbial transglutaminase using Streptomyces sp. polar strains. Romanian Biotechnological Letters. 2010; 15(2): 5197-203.
Serafini-Fracassini D & Del Duca S. Transglutaminases: widespread cross-linking enzymes in plants. Annals of Botany. 2008; 102(2): 145-52.
Duarte L, Matte CR, Bizarro CV & Ayub MAZ. Transglutaminases: part I—origins, sources, and biotechnological characteristics. World Journal of Microbiology and Biotechnology. 2020; 36: 1-18.
Buettner K, Hertel TC & Pietzsch M. Increased thermostability of microbial transglutaminase by combination of several hot spots evolved by random and saturation mutagenesis. Amino Acids. 2012; 42: 987-96.
Nagy V & Szakacs G. Production of transglutaminase by Streptomyces isolates in solid‐state fermentation. Letters in applied microbiology. 2008; 47(2): 122-7.
Yu Y-J, Wu S-C, Chan H-H, Chen Y-C, Chen Z-Y & Yang M-T. Overproduction of soluble recombinant transglutaminase from Streptomyces netropsis in Escherichia coli. Applied microbiology and biotechnology. 2008; 81: 523-32.
Liu X, Yang X, Xie F & Qian S. Cloning of transglutaminase gene from Streptomyces fradiae and its enhanced expression in the original strain. Biotechnology letters. 2006; 28: 1319-25.
Aidaroos H, Du G & Chen J. Microbial fed-batch production of transglutaminase using ammonium sulphate and calcium chloride by Streptomyces hygroscopicus. Biotechnol Bioinf Bioeng. 2011; 1(2): 173-8.
Zotchev SB. Marine actinomycetes as an emerging resource for the drug development pipelines. Journal of biotechnology. 2012; 158(4): 168-75.
Manivasagan P, Venkatesan J, Sivakumar K & Kim S-K. RETRACTED: Marine actinobacterial metabolites: Current status and future perspectives. Microbiological research. 2013; 168(6): 311-32.
Anandan R, Dharumadurai D & Manogaran GP. An introduction to actinobacteria. Actinobacteria-basics and biotechnological applications: IntechOpen, 2016.
Azman A-S, Othman I, Fang C-M, Chan K-G, Goh B-H & Lee L-H. Antibacterial, anticancer and neuroprotective activities of rare Actinobacteria from mangrove forest soils. Indian journal of microbiology. 2017; 57: 177-87.
Salwan R & Sharma V. The role of actinobacteria in the production of industrial enzymes. New and future developments in microbial biotechnology and bioengineering: Elsevier, 2018: 165-77.
Cătălin I, Nicolae B & Gabriela B. Preliminary studies regarding transglutaminase synthesis by polar filamentous bacteria of the genus Streptomyces sp. Innovative Romanian Food Biotechnology. 2009(4): 12-5.
Kanaji T, Ozaki H, Takao T, Kawajiri H, Ide H, Motoki M & et al. Primary structure of microbial transglutaminase from Streptoverticillium sp. strain s-8112. Journal of Biological Chemistry. 1993; 268(16): 11565-72.
Kashiwagi T, Yokoyama K-i, Ishikawa K, Ono K, Ejima D, Matsui H & et al. Crystal Structure of Microbial Transglutaminase fromStreptoverticillium Mobaraense. Journal of Biological Chemistry. 2002; 277(46): 44252-60.
Junqua M, Duran R, Gancet C & Goulas P. Optimization of microbial transglutaminase production using experimental designs. Applied Microbiology and Biotechnology. 1997; 48: 730-734.
Itaya H & Kikuchi Y. Secretion of Streptomyces mobaraensis pro-transglutaminase by coryneform bacteria. Applied microbiology and biotechnology. 2008; 78: 621-5.
Farris M & Olson J. Detection of Actinobacteria cultivated from environmental samples reveals bias in universal primers. Letters in applied microbiology. 2007; 45(4): 376-81.
YokoyAMA K-i, Nakamura N, Seguro K & KuBoTA K. Overproduction of microbial transglutaminase in Escherichia coli, in vitro refolding, and characterization of the refolded form. Bioscience, biotechnology, and biochemistry. 2000; 64(6): 1263-70.
Fuchsbauer HL. Approaching transglutaminase from Streptomyces bacteria over three decades. The FEBS Journal. 2022; 289(16): 4680-703.
Lin Y-S, Chao M-L, Liu C-H, Tseng M & Chu W-S. Cloning of the gene coding for transglutaminase from Streptomyces platensis and its expression in Streptomyces lividans. Process Biochemistry. 2006; 41(3): 519-24.
Macedo JA, Cavallieri ALF, Da Cunha RL & Sato HH. The effect of transglutaminase from Streptomyces sp. CBMAI 837 on the gelation of acidified sodium caseinate. International dairy journal. 2010; 20(10): 673-9.
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Abd-Rabo F, Ei-Dieb S, Abd-EI-Fattah A & Sakr S. Natural state changes of cows' and buffaloes' milk proteins induced by microbial transglutaminase. J Am Sci. 2010; 6(9): 612-20.
Bourneow C, Benjakul S & H-Kittikun A. Hydroxamate-based colorimetric method for direct screening of transglutaminase-producing bacteria. World Journal of Microbiology and Biotechnology. 2012; 28: 2273-7.
Bahrim G, Iancu C, Buţu N & Negoiţă TG. Production of a novel microbial transglutaminase using Streptomyces sp. polar strains. Romanian Biotechnological Letters. 2010; 15(2): 5197-203.
Serafini-Fracassini D & Del Duca S. Transglutaminases: widespread cross-linking enzymes in plants. Annals of Botany. 2008; 102(2): 145-52.
Duarte L, Matte CR, Bizarro CV & Ayub MAZ. Transglutaminases: part I—origins, sources, and biotechnological characteristics. World Journal of Microbiology and Biotechnology. 2020; 36: 1-18.
Buettner K, Hertel TC & Pietzsch M. Increased thermostability of microbial transglutaminase by combination of several hot spots evolved by random and saturation mutagenesis. Amino Acids. 2012; 42: 987-96.
Nagy V & Szakacs G. Production of transglutaminase by Streptomyces isolates in solid‐state fermentation. Letters in applied microbiology. 2008; 47(2): 122-7.
Yu Y-J, Wu S-C, Chan H-H, Chen Y-C, Chen Z-Y & Yang M-T. Overproduction of soluble recombinant transglutaminase from Streptomyces netropsis in Escherichia coli. Applied microbiology and biotechnology. 2008; 81: 523-32.
Liu X, Yang X, Xie F & Qian S. Cloning of transglutaminase gene from Streptomyces fradiae and its enhanced expression in the original strain. Biotechnology letters. 2006; 28: 1319-25.
Aidaroos H, Du G & Chen J. Microbial fed-batch production of transglutaminase using ammonium sulphate and calcium chloride by Streptomyces hygroscopicus. Biotechnol Bioinf Bioeng. 2011; 1(2): 173-8.
Zotchev SB. Marine actinomycetes as an emerging resource for the drug development pipelines. Journal of biotechnology. 2012; 158(4): 168-75.
Manivasagan P, Venkatesan J, Sivakumar K & Kim S-K. RETRACTED: Marine actinobacterial metabolites: Current status and future perspectives. Microbiological research. 2013; 168(6): 311-32.
Anandan R, Dharumadurai D & Manogaran GP. An introduction to actinobacteria. Actinobacteria-basics and biotechnological applications: IntechOpen, 2016.
Azman A-S, Othman I, Fang C-M, Chan K-G, Goh B-H & Lee L-H. Antibacterial, anticancer and neuroprotective activities of rare Actinobacteria from mangrove forest soils. Indian journal of microbiology. 2017; 57: 177-87.
Salwan R & Sharma V. The role of actinobacteria in the production of industrial enzymes. New and future developments in microbial biotechnology and bioengineering: Elsevier, 2018: 165-77.
Cătălin I, Nicolae B & Gabriela B. Preliminary studies regarding transglutaminase synthesis by polar filamentous bacteria of the genus Streptomyces sp. Innovative Romanian Food Biotechnology. 2009(4): 12-5.
Kanaji T, Ozaki H, Takao T, Kawajiri H, Ide H, Motoki M & et al. Primary structure of microbial transglutaminase from Streptoverticillium sp. strain s-8112. Journal of Biological Chemistry. 1993; 268(16): 11565-72.
Kashiwagi T, Yokoyama K-i, Ishikawa K, Ono K, Ejima D, Matsui H & et al. Crystal Structure of Microbial Transglutaminase fromStreptoverticillium Mobaraense. Journal of Biological Chemistry. 2002; 277(46): 44252-60.
Junqua M, Duran R, Gancet C & Goulas P. Optimization of microbial transglutaminase production using experimental designs. Applied Microbiology and Biotechnology. 1997; 48: 730-734.
Itaya H & Kikuchi Y. Secretion of Streptomyces mobaraensis pro-transglutaminase by coryneform bacteria. Applied microbiology and biotechnology. 2008; 78: 621-5.
Farris M & Olson J. Detection of Actinobacteria cultivated from environmental samples reveals bias in universal primers. Letters in applied microbiology. 2007; 45(4): 376-81.
YokoyAMA K-i, Nakamura N, Seguro K & KuBoTA K. Overproduction of microbial transglutaminase in Escherichia coli, in vitro refolding, and characterization of the refolded form. Bioscience, biotechnology, and biochemistry. 2000; 64(6): 1263-70.
Fuchsbauer HL. Approaching transglutaminase from Streptomyces bacteria over three decades. The FEBS Journal. 2022; 289(16): 4680-703.
Lin Y-S, Chao M-L, Liu C-H, Tseng M & Chu W-S. Cloning of the gene coding for transglutaminase from Streptomyces platensis and its expression in Streptomyces lividans. Process Biochemistry. 2006; 41(3): 519-24.
Macedo JA, Cavallieri ALF, Da Cunha RL & Sato HH. The effect of transglutaminase from Streptomyces sp. CBMAI 837 on the gelation of acidified sodium caseinate. International dairy journal. 2010; 20(10): 673-9.
