Identification, cloning, and expression of the novel recombinant xylanase with stable  structure from the sheep rumen microbiota

Ariaeenejad, Shohreh; Maleki, Morteza; Nooshi Nedamani, Safura; Kavousi, Kaveh; Hosseini Salekdeh, Ghasem

Identification, cloning, and expression of the novel recombinant xylanase with stable structure from the sheep rumen microbiota

Document Type : Original Research

Authors

Shohreh Ariaeenejad ¹

Morteza Maleki ¹

Safura Nooshi Nedamani ¹

Kaveh Kavousi ²

Ghasem Hosseini salekdeh ¹

¹ Department of Systems and Synthetic Biology, Agricultural Biotechnology Research Institute of Iran (ABRII), Agricultural Research Education and Extension Organization (AREEO), Karaj, Iran.

² Laboratory of Complex Biological Systems and Bioinformatics (CBB), Institute of Biochemistry and Biophysics (IBB), University of Tehran, Tehran, Iran

Abstract

Enzymes play an essential role in catalyzing the reactions for multiple industrial applications. One of these critical industries with a worldwide application is paper and pulp, which is cost-effective in increasing attention. Xylanases are potential enzymes that proved their abilities in a broad range of applications, specifically in the paper and pulp industry as a biobleaching agent and dye removal biocatalyst. In these decades, the production of novel enzymes from natural sources is conceivable, especially with applying the culture-independent method of metagenome. This practical approach provides the opportunity to identify the novel enzymes from uncultivable microbial diversities. Concerning the importance of the thermostable enzymes for industrial applications and their better action in harsh conditions, this study aimed to identify novel thermostable xylanase from metagenomic data of sheep rumen by applying the in-silico screening. The thermostable xylanase was extracted from the ruminal DNA and after cloning and expression named PersiXyn5. The enzymeschr('39') kinetic parameters, including Km, Vmax, and its specific activity, were examined. The enzyme was optimally active at 80 and pH 8 and could retain 58% of its maximum activity after 2h of incubation at 90 . The thermostable, alkali PersiXyn5 was an efficient enzyme in the paper industry and poultry feed and fuel applications.

Keywords

Novel Xylanase

In-silico screening

Metagenome

thermostability

Subjects

Microbial biotechnology

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