Cloning of the xylanase gene from soil Streptomyces into Escherichia coli for the poultry industry application

PDF (English)
DOI : https://doi.org/10.12681/jhvms.29390
M BAHRAMINIA
Department of Microbiology, Faculty of Sciences, Central Branch, Islamic Azad University, Tehran, Iran
F MORADMAND
Department of Microbiology, Faculty of Sciences, Saveh Branch, Islamic Azad University, Saveh, Iran
K AMINI
Department of Microbiology, Faculty of Sciences, Saveh Branch, Islamic Azad University, Saveh, Iran
Résumé

Streptomyces are gram-positive aerobic strains that are isolated from soil, water, sediments, and various sources. The bacteria are capable of producing secondary metabolites, such as enzymes that sometimes play unique functional roles in industry, and are one of the important bio-control agents. This study aimed to isolate and clone thexylanase gene from soil Streptomyces. Soil samples were collected from Markazi Province, Iranafter specific biochemical examinations, isolation of bacteria, and DNA extraction. PCR was then performed to identify the strains containing the xylanase gene. The gene from the positive strains was cloned into an E.coli host-vector by TA cloning technique and finally, the expression of genes in E. coli origami was measured by Real-Time PCR technique. ClustalX and Mega5 software were used to draw the phylogenetic tree. A total of twelve Streptomyces isolates were identified from the soil samples. Among all the isolates, threeones had the xylanase gene. After cloning the xylanase genes, the cloned strains were isolated. To confirm the DNA cloning, Real-Time PCR was performed, and finally, the PCR product where sequenced. In this study, Streptomyces was identified as a native strain for the expression of xylanase after generating recombinant plasmid and TA cloning. It can be stated that cloning of the xylanase gene from soil Streptomyces in E. coli can be used in the poultry industry.

Article Details
  • Rubrique
  • Research Articles
Creative Commons License

Ce travail est disponible sous licence Creative Commons Attribution - Pas d’Utilisation Commerciale 4.0 International.

Authors who publish with this journal agree to the following terms:

· Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution Non-Commercial License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.

· Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g. post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.

· Authors are permitted and encouraged to post their work online (preferably in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work.

Téléchargements
Les données relatives au téléchargement ne sont pas encore disponibles.
Références
Ahmed S, Riaz S, Jamil A (2009) Molecular cloning of fungal xylanases: an overview. Appl MicrobiolBiotechnol 84: 19-35.
Akyol I, Comlekcioglu U, Kar B, Ekinci MS, Ozkose E (2009) Cloning of a xylanase gene xyn2A from rumen fungus Neocallimastix sp. GMLF2 in Escherichia coli and its partial characterization. Biologia 64: 664-670.
Emam MTH, Mohamed KA, Badawy FMI, and Ibrahim SA (2016) Cloning, sequencing and expression of the xylanase genefrom Bacillus pumilus GH in Escherichia coli. Int J ChemTech Res9(12): 114-124.
Guo B, Chen XL, Sun CY, Zhou BC, Zhang YZ (2009) Gene cloning, expression and characterization of a new old-active and salt-tolerant endo-β-1,4-xylanase from marine Glaciecola mesophila KMM 241. Appl Microbiol Biotechnol 84: 1107-1115.
Goswami GK, Krishnamohan M, Nain V, Aggarwal C, Ramesh B (2014) Cloning and heterologous expression of cellulose free thermostable xylanase from Bacillus brevis. Springer Plus 3:20-26.
Hwang IT, Lim HK, Song HY, Cho SJ, Chang JS, Park NJ (2010) Cloning and characterization of a xylanase, KRICT PX1 from the strain Paenibacillus sp. HPL-001. Biotech Adv 28: 594-601.
Jeya M, Thiagarajan S, Lee JK, Gunasekaran P (2009) Cloning sequencing and expresstion of GHxylanase geneAspergillus fumigatus MKU1 in pichia pastoris. J Biosci Bioeng 108: 24-29.
Kishishita S, Yoshimi M, Fujii T, Taylor LE, Decker SR, Ishikawa K, Inoue H (2014) Celluloseinducible xylanase Xyl10A from Acremonium cellulolyticus: Purification, cloning and homologous expression. Protein Expr Purif 94: 40-45.
Li N, Meng K, Wang Y, Shi P, Luo H, Bai Y, Yang P, Yao B (2008) Cloning, expression, and characterization of a new xylanase with broad temperature adaptability from Streptomyces sp. S9. Appl Microbiol Biotechnol 80:231-240.
Mirzaie S, Zaghari M, Aminzadeh S, Shivazad M (2012)The effects of non-starch polysaccharidescontent of wheat and xylanase supplementation onthe intestinal amylase, aminopeptidase and lipaseactivities, ileal viscosity and fat digestibility inlayer diet. Iran J Biotech 10: 208-214.
Polizeli MLTM, Rizzatti ACS, Monti R, Terenzi HF,Jorge JA, Amorim DS (2005) Xylanases fromfungi: properties and industrial applications. ApplMicrobiol Biotechnol 67: 577-591.
Verma D, KawarabayasiY, Miyazaki K, Satyanarayana T (2013) Cloning, Expression and Characteristics of a Novel Alkalistable and Thermostable Xylanase Encoding Gene (Mxyl) Retrieved from Compost- Soil Metagenome. PLoS ONE 8(1): e52459. doi:10.1371/journal.pone.0052459.
Sevillano L, Vijgenboom E, van Wezel GP, Santamara RI (2016) New approaches to achieve high level enzyme production in Streptomyces lividans. Microb Cell Fact 15:28. Doi: 10.1186/s12934-016-0425-7.
Subramaniyan S, Sandhia GS, Prema P (2001) Control of xylanase production without protease activity in Bacillus sp. by selection of nitrogen source. Biotechnol Lett 23: 369-371.
Wang G, Luo H, Wang Y, Huang H, Shi P, Yang P,Meng K, Bai Y, Yao B (2011) A novel cold-activexylanase gene from the environmental DNA ofgoat rumen contents: direct cloning, expression andenzyme characterization. Bioresour Technol 102:3330-3336.
Articles les plus lus par le même auteur ou la même autrice