Aspergillus niger production of pectinase and α-galactosidase for enzymatic soy processing.

Soybean is a most promising sustainable protein source for feed and food to help meet the protein demand of the rapidly rising global population. To enrich soy protein, the environment-friendly enzymatic processing requires multiple carbohydrases including cellulase, xylanase, pectinase, α-galactosidase and sucrase. Besides enriched protein, the processing adds value by generating monosaccharides that are ready feedstock for biofuel/bioproducts. Aspergillus could produce the required carbohydrases, but with deficient pectinase and α-galactosidase. Here we address this critical technological gap by focused evaluation of the suboptimal productivity of pectinase and α-galactosidase. A carbohydrases-productive strain A. niger (NRRL 322) was used with soybean hull as inducing substrate. Temperatures at 20 °C, 25 °C and 30 °C were found to affect cell growth on sucrose with an Arrhenius-law activation energy of 28.7 kcal/mol. The 30 °C promoted the fastest cell growth (doubling time = 2.1 h) and earliest enzyme production, but it gave lower final enzyme yield due to earlier carbon-source exhaustion. The 25 °C gave the highest enzyme yield. pH conditions also strongly affected enzyme production. Fermentations made with initial pH of 6 or 7 were most productive, e.g., giving 1.9- to 2.3-fold higher pectinase and 2.2- to 2.3-fold higher α-galactosidase after 72 h, compared to the fermentation with a constant pH 4. Further, pH must be kept above 2.6 to avoid limitation in pectinase production and, in the later substrate-limiting stage, kept below 5.5 to avoid pectinase degradation. α-Galactosidase production always followed the pectinase production with a 16-24 h lag; presumably, the former relied on pectin hydrolysis for inducers generation. Optimal enzyme production requires controlling the transient availability of inducers.

High monomeric sugar yields from enzymatic hydrolysis of soybean meal and effects of mild heat pretreatments with chelators.

Defatted soybean meal has 30-35% oligo-/polymeric carbohydrates and approximately 50% proteins. Enzymatic carbohydrate monomerization enables easy separation to enrich protein content, reduces indigestibility concerns, and facilitates use of carbohydrate as fermentation feedstock. Among soybean carbohydrates, pectin and glucan are more recalcitrant to hydrolyze. To destabilize Ca2+-bridged junctures in pectin, effects of 3 chelators ethylenediaminetetraacetic acid (EDTA), sodium hexametaphosphate (HMP) and citric acid under 2-h 90 °C pretreatments were investigated here. Citric acid was the most effective while EDTA decreased enzymatic hydrolysis. In a 3-factor 2-level factorial study, heat (90 °C, 2 h) and citric acid (10 g/L) pretreatments and cellulase supplementation (10 FPU/g) were found to increase yields of all monosaccharides, to 86.8 ±â€¯5.2% glucose, 98.1 ±â€¯1.6% xylose, 87.5 ±â€¯5.2% galactose, 83.6 ±â€¯1.6% arabinose, and 91.4 ±â€¯3.1% fructose + mannose. The largest percentage improvements were for arabinose (382%), mannose (113%) and glucose (51%). Achieving high monosaccharide yields greatly increases value of soybean carbohydrate as fermentation feedstock.

Production of arabitol from enzymatic hydrolysate of soybean flour by Debaryomyces hansenii fermentation.

Arabitol is a low-calorie sugar alcohol with anti-cariogenic properties. Enzymatic hydrolysate of soybean flour is a new renewable biorefinery feedstock containing hexose, pentose, and organic nitrogen sources. Arabitol production by Debaryomyces hansenii using soybean flour hydrolysate was investigated. Effects of medium composition, operating conditions, and culture stage (growing or stationary phase) were studied. Production was also compared at different culture volumes to understand the effect of dissolved oxygen concentration (DO). Main factors examined for medium composition effects were the carbon to nitrogen concentration ratio (C/N), inorganic (ammonium) to organic nitrogen ratio (I/O-N), and sugar composition. Arabitol yield increased with increasing C/N ratio and a high I/O-N (0.8-1.0), suggesting higher yield at stationary phase of low pH (3.5-4.5). Catabolite repression was observed, with the following order of consumption: glucose > fructose > galactose > xylose > arabinose. Arabitol production also favored hexoses and, among hexoses, glucose. DO condition was of critical importance to arabitol production and cell metabolism. The yeast consumed pentoses (xylose and arabinose) only at more favorable DO conditions. Finally, arabitol was produced in fermentors using mixed hydrolysates of soy flour and hulls. The process gave an arabitol yield of 54%, volumetric productivity of 0.90 g/L-h, and specific productivity of 0.031 g/g-h.