Development and validation of a medium for recombinant endo-β-1,4- xylanase production by Kluyveromyces lactis using a statistical experimental design

Siti Fatimah Zaharah Mohamad Fuzi, Nor Muhammad Mahadi, Jamaliah Md Jahim, Abdul Munir Abd. Murad, Farah Diba Abu Bakar, Mazura Jusoh, Roshanida A. Rahman, Rosli Md Illias

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Kluyveromyces lactis is an excellent host for a high cell density culture, which allows high expression levels of recombinant enzymes. Nutrient composition and culture conditions affect the secretion, production level and stability of the recombinant host. Therefore, it is technologically important to formulate a medium that stimulates high cell density and enhances the desired enzyme production using K. lactis GG799. In this study, six media were initially compared, and a Plackett-Burman experimental design was employed to screen for important components and trace elements. Nitrogen sources such as ammonium sulfate and free amino acid (casamino acid) as well as compounds like MgSO 4·7H 2O, Na 2SO 4, ZnSO 4·6H 2O, MnSO 4·4H 2O and KH 2PO 4 affected biomass concentrations (5.67 g/l) and recombinant endo-β-1,4-xylanase (Xyn2) production (49.73 U/ml). Optimum productivity was obtained at shorter incubation times (i.e., 6 h), making the medium suitable for use when seeking efficient production. Expression of recombinant Xyn2 by K. lactis GG799 in the designed medium resulted in satisfactory recombinant Xyn2 volumetric productivity (v p) at 8.29 U/ml/h. When compared to the rich, non-selective YPD medium, the designed medium improved biomass output and recombinant Xyn2 production in K. lactis GG799 by approximately 9 and 22%, respectively.

Original languageEnglish
Pages (from-to)283-292
Number of pages10
JournalAnnals of Microbiology
Volume62
Issue number1
DOIs
Publication statusPublished - Mar 2012

Fingerprint

Kluyveromyces
Biomass
Research Design
Cell Count
Ammonium Sulfate
Trace Elements
Enzymes
Nitrogen
Cell Culture Techniques
Amino Acids
Food
casamino acids

Keywords

  • Kluyveromyces lactis GG799
  • Medium development
  • Plackett-Burman
  • Recombinant xylanase

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology

Cite this

Development and validation of a medium for recombinant endo-β-1,4- xylanase production by Kluyveromyces lactis using a statistical experimental design. / Fuzi, Siti Fatimah Zaharah Mohamad; Mahadi, Nor Muhammad; Md Jahim, Jamaliah; Abd. Murad, Abdul Munir; Abu Bakar, Farah Diba; Jusoh, Mazura; Rahman, Roshanida A.; Illias, Rosli Md.

In: Annals of Microbiology, Vol. 62, No. 1, 03.2012, p. 283-292.

Research output: Contribution to journalArticle

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