Expression of multi-functional cellulase gene mfc in Coprinus cinereus under control of different basidiomycete promoters.
Multi-functional cellulase gene mfc was expressed in Coprinus cinereus under naturally non-inductive conditions using three heterologous promoters. Endo-beta-1,4-glucanase expression was achieved in solid and liquid media with promoter sequences from the Lentinula edodesgpd gene, the Flammulina velutipes gpd gene and the Volvariella volvaceagpd gene. As measured by enzyme activity in liquid cultures, a 613-bp gpd promoter fragment from L. edodes was most efficient, followed by a 752-bp gpd fragment from F. velutipes. The V. volvacea gpd promoter sequence was less active, in comparison. Irrespective of the promoter used, enzymatic activities increase 34-fold for highly active transformants and 29-fold for less active one by using cellulase-inducing medium. The highest activities of endo-beta-1,4-glucanase (34.234 U/ml) and endo-beta-1,4-xylanase (263.695 U/ml) were reached by using the L. edodesgpd promoter.
Department of Bioengineering, College of Food Science, South China Agricultural University, Tianhe, Guangzhou, China.
Shujie
S
Cheng
Peizhou
P
Yang
Liqiong
L
Guo
Junfang
J
Lin
Nannan
N
Lou
D015139:Blotting, Southern; D002480:Cellulase / Q000096:biosynthesis / Q000235:genetics*; D003224:Congo Red; D003302:Coprinus / Q000201:enzymology* / Q000235:genetics*; D003470:Culture Media; D004587:Electrophoresis, Agar Gel; D005656:Fungal Proteins / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic; D014170:Transformation, Genetic
D003470:Culture Media; D005656:Fungal Proteins; D012333:RNA, Messenger; D003224:Congo Red; D002480:Cellulase