title: Recombinant Candida utilis for the production of biotin.
abstract: Biotin is an important nutritional supplement but is difficult to manufacture effectively. Here we present a trial of biotin production using the food yeast Candida utilis. In this system, we cloned the C. utilis biotin synthase (BIO2) gene, the gene of the rate-limiting enzyme for biotin biosynthesis, and assembled it under the control of a strong promoter. A series of plasmids were constructed to direct the integration of the BIO2 gene, either high-copy integration with 18S rDNA fragment or low-copy integration with URA3 or HIS3 fragment. The BIO2 gene can be successfully integrated into the C. utilis chromosome and can drive biotin production using these plasmids. The biotin yield in this system can reach 100-fold above the endogenous level in a small-scale culture. Although the biotin production is not stable if the selection pressure is removed, this system has the potential to produce biotin-rich feed or food additives directly without the requirement of further purification.
authors.affiliation: Institute of Biochemistry, Kaohsiung Medical University, Taiwan, ROC.
authors.forename: Yi-Ren
authors.initials: YR
authors.lastname: Hong
authors.forename: Ya-Lei
authors.initials: YL
authors.lastname: Chen
authors.forename: Lynn
authors.initials: L
authors.lastname: Farh
authors.forename: Wen-Jen
authors.initials: WJ
authors.lastname: Yang
authors.forename: Chen-Hua
authors.initials: CH
authors.lastname: Liao
authors.forename: David
authors.initials: D
authors.lastname: Shiuan
mesh_terms: D001710:Biotin / Q000096:biosynthesis* / Q000235:genetics; D002175:Candida / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D007218:Industrial Microbiology / Q000379:methods*; D016297:Mutagenesis, Site-Directed; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D011995:Recombination, Genetic; D013466:Sulfurtransferases / Q000235:genetics / Q000378:metabolism*; D014170:Transformation, Genetic
chemical_list: D004271:DNA, Fungal; D011994:Recombinant Proteins; D001710:Biotin; D013466:Sulfurtransferases; C057985:biotin synthetase