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Cloning, Expression and Characterization of The Thermophilic Xylanase gene (xyn11) in Pichia pastoris KM71

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ABSTRACT

Xylanases are hydrolytic enzymes which randomly cleave the β 1,4 backbone of
the complex plant cell wall of polysaccharide in xylan. In this study, xyn11 gene
that encoding thermophilic xylanase from bagasse metagenome was cloned into E.
coli DH5α and transformation in Pichia pastoris KM71 furthermore characterized
by biochemical properties consists of optimum pH, temperature, thermostability,
and substrate specificity. In this study demonstrated xyn11 gene from bagasse
metagenome was successfully cloned and expressed in methylotrophic yeast P.
pastoris KM71 as secreted form (9 mg/L production media). Recombinant Xyn11
protein showed specific activity toward only hemicelluloses with 4652.715 U/mg
protein and 179.116 U/mg protein on beechwood xylan and arabinoxylan,
respectively. However, Xyn11 lacks hydrolytic activity against different forms of
cellulose (Avicel PH101 microcrystalline cellulose, phosphoric acid swollen
cellulose and carboxymethylcellulose). Xyn11 showed the highest activity in
sodium phosphate buffer pH 6.0 at 80C. However, Xyn11 remained only 60%
relative activity when it was incubated at 40C for 15 min.
Keywords: Xylanase, Cloning, Epression gene, Biochemical characterization,
Pichia pastoris KM71

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Ketersediaan

1256B18IV

1256 B 18

Perpustakaan FSM Undip (Referensi)

Tersedia

Informasi Detail

Judul Seri

BIOLOGI

No. Panggil

1256 B 18

Penerbit

: ., 2018

Deskripsi Fisik

-

Bahasa

Indonesia

ISBN/ISSN

-

Klasifikasi

004

Tipe Isi

-

Tipe Media

-

Tipe Pembawa

-

Edisi

-

Subjek

-

Info Detail Spesifik

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Pernyataan Tanggungjawab

Salma Fuadiyah

Versi lain/terkait

Lampiran Berkas

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