Production of Xylanase by Recombinant Bacillus subtilis DB104 Cultivated in Agroindustrial Waste Medium

  • Is Helianti Center for Bioindustrial Technology, Agency for Assessment and Application of Technology (BPPT), LAPTIAB-BPPT, PUSPIPTEK Serpong, Tangerang Selatan, Indonesia
  • Maria Ulfah Center for Bioindustrial Technology, Agency for Assessment and Application of Technology (BPPT), LAPTIAB-BPPT, PUSPIPTEK Serpong, Tangerang Selatan, Indonesia
  • Niknik Nurhayati Center for Bioindustrial Technology, Agency for Assessment and Application of Technology (BPPT), LAPTIAB-BPPT, PUSPIPTEK Serpong, Tangerang Selatan, Indonesia
  • Dadang Suhendar Center for Bioindustrial Technology, Agency for Assessment and Application of Technology (BPPT), LAPTIAB-BPPT, PUSPIPTEK Serpong, Tangerang Selatan, Indonesia
  • Anita Kusuma Finalissari Center for Bioindustrial Technology, Agency for Assessment and Application of Technology (BPPT), LAPTIAB-BPPT, PUSPIPTEK Serpong, Tangerang Selatan, Indonesia Department of Agricultural Product Technology, Faculty of Agriculture Technology, University of Brawijaya, Malang, Indonesia
  • Agustin Krisna Wardani Department of Agricultural Product Technology, Faculty of Agriculture Technology, University of Brawijaya, Malang, Indonesia
Keywords: corncobs, cost-effective medium, endoxylanase, recombinant Bacillus subtilis DB104, tofu liquid waste, xylooligosaccharides

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.

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Published
2017-04-14
Section
Articles