Han Sung Ok, Cho Hee-Yeon, Yukawa Hideaki, Inui Masayuki, Doi Roy H
Section of Molecular and Cellular Biology, University of California, Davis, Davis, California 95616, USA.
J Bacteriol. 2004 Jul;186(13):4218-27. doi: 10.1128/JB.186.13.4218-4227.2004.
Cellulosomes and noncellulosomal (hemi)cellulolytic enzymes are produced by Clostridium cellulovorans to degrade plant cell walls. To understand their synergistic relationship, changes in mRNA and protein expression in cellulosomes and noncellulosomal (hemi)cellulolytic enzymes (hereafter called noncellulosomal enzymes) of cultures grown on cellobiose, cellulose, pectin, xylan, and corn fiber or mixtures thereof were examined. Cellulase expression, favored particularly by the presence of Avicel, was found with all substrates. Comparison of cellulosome and noncellulosomal enzymes showed that expression profiles were strongly affected by the carbon source. High xylanase or pectate lyase expression was observed when C. cellulovorans was grown on xylan or pectin, respectively. Mixed carbon substrates (cellulose-pectin-xylan mixture or corn fiber) induced a wider variety of enzymes than a single carbon source, such as cellobiose, pectin, or xylan. Cellulosomal proteome profiles were more affected by the carbon source than the noncellulosomal enzymes. Transcription and protein analyses revealed that cellulosomes and noncellulosomal enzymes were expressed simultaneously on mixed carbon sources, but their degree of inducibility varied when the substrate was either cellulose or cellobiose. Cellulosomes and noncellulosomal enzymes had synergistic activity on various carbon substrates. These results indicated that expression of plant cell wall-degrading enzymes is highly influenced by the available carbon source and that synergy between cellulosomes and noncellulosomal enzymes contribute to plant cell wall degradation.
嗜纤维梭菌可产生纤维小体和非纤维小体(半)纤维素分解酶,用于降解植物细胞壁。为了解它们的协同关系,研究了在纤维二糖、纤维素、果胶、木聚糖、玉米纤维或其混合物上生长的培养物中,纤维小体和非纤维小体(半)纤维素分解酶(以下简称非纤维小体酶)的mRNA和蛋白质表达变化。发现所有底物都能促进纤维素酶的表达,尤其是微晶纤维素的存在。纤维小体酶和非纤维小体酶的比较表明,表达谱受碳源的强烈影响。当嗜纤维梭菌分别在木聚糖或果胶上生长时,观察到高木聚糖酶或果胶酸裂解酶表达。混合碳底物(纤维素-果胶-木聚糖混合物或玉米纤维)比单一碳源(如纤维二糖、果胶或木聚糖)诱导产生更多种类的酶。纤维小体蛋白质组谱比非纤维小体酶受碳源的影响更大。转录和蛋白质分析表明,在混合碳源上,纤维小体和非纤维小体酶同时表达,但当底物为纤维素或纤维二糖时,它们的诱导程度有所不同。纤维小体和非纤维小体酶在各种碳底物上具有协同活性。这些结果表明,植物细胞壁降解酶的表达受可用碳源的高度影响,并且纤维小体和非纤维小体酶之间的协同作用有助于植物细胞壁的降解。