Enhanced Nutritional Composition of Steam-Exploded Cotton Stalk Through Microbial-Enzyme Synergism Solid-State Fermentation

Due to its high content of lignocellulose, cotton stalk is difficult to degrade naturally and utilize effectively, so it is often regarded as waste. In this study, the effects of Pleurotus ostreatus XH005, Lactiplantibacillus plantarum LP-2, and cellulase enzyme on the cotton stalk substrate under aerobic solid-state fermentation (SSF) conditions were investigated, and the metabolites were analyzed to identify potential functional compounds in the cotton-stalk-fermented feed. Preliminary optimization results obtained through single-factor experiments were as follows: fermentation time 14 days, XH005 inoculum size 8.00% (v/m), material-to-water ratio 1:0.50 (v/m), LP-2 inoculum size 2.00% (v/m), and cellulase addition 0.60% (m/m). Based on these single-factor experimental results, XH005 inoculum size, LP-2 inoculum size, material-to-water ratio, and cellulase addition were selected as independent variables. Through response surface methodology (RSM) optimization experiments, 29 experimental groups were designed. Subsequently, based on Box–Behnken analysis of variance (ANOVA) of lignin and cellulose content, along with contour and response surface plots, the optimal aerobic solid-state fermentation parameters were determined as follows: fermentation time 14 days, XH005 inoculum: 7.00% (v/m), material-to-water ratio: 1:0.55 (v/m), LP-2 inoculum: 2.00% (v/m), and cellulase enzyme addition: 0.65% (m/m). Results showed that compared with the control group (CK), the optimized group exhibited a 27.65% increase in lignin degradation rate and a 47.14% increase in cellulose degradation rate. Crude protein (CP) content increased significantly, while crude fiber (CF), detergent fiber and mycotoxin contents decreased significantly. Non-targeted metabolic analysis indicated that adding cellulase and inoculating Pleurotus ostreatus XH005 and Lactiplantibacillus plantarum LP-2 in aerobic SSF of cotton straw feed produced functionally active substances such as kaempferol (C343), carvone (C709) and trilobatin (C604). Therefore, this study demonstrates that microbial-enzyme co-action SSF significantly enhances the nutritional composition of cotton stalk hydrolysate. Furthermore, this hydrolysate is suitable for the production of functional compounds, endowing the fermented feed with health-promoting properties and enhancing the utilization of cotton processing byproducts in the feed industry.