Department Biologie, Molekulare Mykologie, Heinrich-Heine-Universität, Universitätsstrasse 1/26.12.01, 40225, Düsseldorf, Germany.
Appl Microbiol Biotechnol. 2015 Oct;99(19):8055-64. doi: 10.1007/s00253-015-6703-1. Epub 2015 Jun 9.
The fodder yeast Candida utilis is able to use xylose mono- and oligomers as sources of carbon but not the abundant polymer xylan. C. utilis transformants producing the Penicillium simplicissimum xylanase XynA were constructed using expression vectors encoding fusions of the Saccharomyces cerevisiae Mfα1 pre-pro secretion leader to XynA. The Mfα1-XynA fusion was efficiently processed in transformants and XynA was secreted almost quantitatively into the culture medium. Secreted XynA was enzymatically active and allowed transformants to grow on xylan as the sole carbon source. Addition of a second expression unit for the heterologous green fluorescent protein (GFP) generated C. utilis transformants, which showed intracellular GFP fluorescence during growth on xylan. The results suggest that xylanase-producing C. utilis is suited as a cost-effective host organism for heterologous protein production and for other biotechnical applications.
饲料酵母 Candida utilis 能够将木糖单糖和寡糖用作碳源,但不能利用丰富的聚合物木聚糖。使用表达载体构建了产生青霉菌 simplicissimum 木聚糖酶 XynA 的 C. utilis 转化体,该载体编码了 Saccharomyces cerevisiae Mfα1 前导肽到 XynA 的融合。Mfα1-XynA 融合在转化体中得到有效处理,XynA 几乎全部分泌到培养基中。分泌的 XynA 具有酶活性,使转化体能够以木聚糖作为唯一碳源生长。添加第二个异源绿色荧光蛋白 (GFP) 的表达单元,产生了 C. utilis 转化体,在木聚糖上生长时表现出细胞内 GFP 荧光。结果表明,产木聚糖酶的 C. utilis 适合作为异源蛋白生产和其他生物技术应用的经济高效的宿主生物。