Proteinase and glycoside hydrolase production is enhanced in solid-state fermentation by manipulating the carbon and nitrogen fluxes in Aspergillus oryzae.

Soy sauce materials of soybean meal and wheat bran were evaluated in solid-state (koji) fermentation (SSF) and submerged fermentation (SmF) by Aspergillus oryzae. Proteinase production in SSF (2331 ± 39 U g-1) was about 4.9 times higher than that in SmF (477 ± 13 U g-1), and glycoside hydrolase was approximately 2 times higher in SSF than that in SmF. In addition, protein expression of iTRAQ analysis deepens our understanding of the secreting mechanism. Abundant proteinases (dipeptidase, dipeptidyl aminopeptidase, puromycin-sensitive aminopeptidase, Xaa-pro aminopeptidase, neutral protease 2 and leucine aminopeptidase 2), along with the glycoside hydrolase (glycoamylase, glucosidase and β-xylanase) were secreted at the late stage of SSF, but tripeptidyl peptidase sed 2 was proposed as an indispensable protease in SmF or the early stage of SSF. Several metabolites associated with the carbon flux and amino acid biosynthesis were proved to be regulated by the proteinase and glycoside hydrolase production.