IS HELIANTI
Center for Bioindustrial Technology, Laboratorium of Bioindustrial Technology, LAPTIAB BPPT Puspiptek -Serpong

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PRODUCTION OF XYLANASE BY RECOMBINANT BACILLUS SUBTILIS DB104 CULTIVATED IN AGROINDUSTRIAL WASTE MEDIUM Helianti, Is; Ulfah, Maria; Nurhayati, Niknik; Suhendar, Dadang; Finalissari, Anita Kusuma; Wardani, Agustin Krisna
HAYATI Journal of Biosciences Vol. 23 No. 3 (2016): July 2016
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1064.942 KB) | DOI: 10.4308/hjb.23.3.125

Abstract

A recombinant Bacillus subtilis DB104 strain harbouring recombinant plasmid pSKE194 containing an Open Reading Frame (ORF) of endoxylanase and its indigenous promoter from the wild-type B. subtilis AQ1 strain was constructed. This recombinant B. subtilisDB104 strain had higher endoxylanase activity than the nonrecombinant B. subtilisDB104 strain in standard media, such as Luria Bertani (LB) and LB with xylan. The agroindustrial wastes corncobs and tofu liquid waste were chosen as cost-effective carbon and nitrogen sources, respectively, to test the economics of xylanase production using the recombinant B. subtilis DB104 at a larger scale. Submerged fermentation using a 4.5 L working volume fermentor with tofu liquid waste and 4% corncobs produced maximum xylanase activity of 1296 ± 1.2 U/mg (601.7 ± 0.6 U/mL) after 48-hour fermentation at 37°C with 150 rpm agitation; this is more than twofold higher than the activity produced in an Erlenmeyer flask. This is the first report of high xylanase activity produced from recombinant B. subtilis using inexpensive medium. During fermentation, the xylanase degrades corncobs into xylooligosaccharides, showing its potential as an enzyme feed additive or in xylooligosaccharide production.
KLONING DAN SEKUENSING GEN XILANASE DENGAN PRODUK GEN BERUKURAN 30 KDA DARI BACILLUS HALODURANS CM1 PADA ESCHERICHIA COLI DH5α Safirah, Dearesty; Helianti, Is; Kusumaningrum, Hermin Pancasakti; Budiharjo, Anto
Bioma : Berkala Ilmiah Biologi Vol. 18, No.2, Tahun 2016
Publisher : Departemen Biologi, Fakultas Sains dan Matematika, Universitas Diponegoro

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (98.71 KB) | DOI: 10.14710/bioma.18.2.167-172

Abstract

The paper industry contributed the environment pollution due to chlor substances. Utilization of alkalothermophilic xylanase enzyme as a biocatalyst in the production of paper may become an environmentally friendly biobleaching alternative. Bacillus halodurans CM1 produces xilanase enzyme that had optimal activity at pH 9 and temperature 70°C. Previous study showed that this CM1 strains has several xilanase genes. The cloning of one of these alkalothermophiic xylanase (alkxyn) gene has been already conducted. This study aimed to clone alkxyn gene that encode alkalothermophilic xylanase enzyme from B. halodurans CM1 into Escherichia coli DH5?. Amplification of alkxyn has been carried out using primers for amplification xylanase 30 kDa. The alkxyn gene fragment was inserted into pGEM-T Easy vector and then transformed into E. coli DH5?. The results showed that the recombinant of E. coli DH5? harboring alkxyn gene from B. halodurans CM1 has been obtained. The sequences analysist based on BLAST showed that alkxyn fragment has homology (99%) with the alkaliphilic xylanase gene from Bacillus sp. 31 which encodes alkaliphilic xilanase (Genebank assession number: JF912895.1). Keywords: cloning, Bacillus halodurans CM1, xylanase, alkalothermophilic.
DIRECT CLONING OF A XYLANASE GENE FROM PAWAN-RIAU HOT SPRING HELIANTI, IS
HAYATI Journal of Biosciences Vol. 14 No. 2 (2007): June 2007
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (137.564 KB) | DOI: 10.4308/hjb.14.2.54

Abstract

A functional gene containing an Open Reading Frame (ORF) encoding a ?-1, 4-endoxylanase glycosyl hydrolase family 11 was cloned directly using metagenomic PCR-cloning method from Pawan Hot Spring sample in Riau. The gene consisted of 642 nucleotides, encoded for 213 amino acids. The amino acid sequence analysis using BLAST showed that the gene has high homology (93%) with xylanase gene from Bacillus subtilis. The gene showed its function when it was subcloned into an expression vector and overexpressed in E. coli. The crude extract of the recombinant enzyme had activity for 170 U/ml at 50 oC. The result of this work showed that metagenomic approach was a powerful short cut method to obtain recombinant biocatalyst that was useful for industrial application. Key words: ?-1, 4-endoxylanase, metagenomic DNA, Pawan-Riau hot-spring