Agricultural straws, such as rice straw, wheat straw, and corn straw, are produced abundantly every year but not utilized efficiently in China. An experiment was conducted to determine the effects of recombinant xylanase on ruminal fermentation and microbial community structure in in vitro incubation of these straws. The recombinant xylanase from Lentinula edodes (rLeXyn11A) was produced in Pichia pastoris. The optimal temperature and pH for rLeXyn11A were 40∘C and 4.0, respectively. The rLeXyn11A featured resistance to high temperature and showed broad temperature adaptability (>50% of the maximum activity at 20–80∘C). Supplemental rLeXyn11A enhanced the hydrolysis of three agricultural straws. After in vitro ruminal incubation, regardless of agricultural straws, the fiber digestibility, acetate concentration, total volatile fatty acids (VFAs) production, and fermentation liquid microbial protein were increased by rLeXyn11A. Supplemental rLeXyn11A increased the ammonia-N concentration for corn straw and rice straw. High throughput sequencing and real-time PCR data showed that the effects of rLeXyn11A on ruminal microbial community depended on the fermentation substrates. With rice straw, rLeXyn11A increased the relative abundance of fibrolytic bacteria including Firmicutes, Desulfovibrio, Ruminococcaceae and its some genus, and Fibrobacter succinogenes. With wheat straw, rLeXyn11A increased the relative abundance of Ruminococcus_1 and its three representative species F. succinogenes, Ruminococcus flavefaciens, Ruminococcus albus. With corn straw, the fibrolytic bacteria Firmicutes, Christensenellaceae_R_7_group, Saccharofermentans, and Desulfovibrio were increased by rLeXyn11A. This study demonstrates that rLeXyn11A could enhance in vitro ruminal digestion and fermentation of agricultural straws, showing the potential of rLeXyn11A for improving the utilization of agricultural straws in ruminants.

Rape straw cannot be efficiently degraded and utilized by ruminants due to its severe lignification and complex cross-linked structure between fiber and lignin. The laccases can catalyze the inter-unit bond cleavage in lignin substrates. Therefore, this study investigated the recombinant laccase from Lentinula edodes (LeLac) and its application in degrading rape straw. The LeLac was expressed using Pichia pastoris. It had the maximum activity at 60°C and pH 3.0 using ABTS as substrate and at 50°C and pH 4.0 using o-tolidine as substrate. The LeLac exhibited preferential oxidation of ABTS and featured resistance to high temperature, but relatively poor thermal stability. The LeLac activity could be strengthened by Cu²⁺ in dose-dependent manners. The LeLac could tolerate 15% of ethanol and methanol. The optimal pH for the lignin degradation of rape straw acid detergent fiber (ADF) by LeLac was 4.0. The LeLac could improve the cellulose enzymolysis of rape straw ADF by degrading its lignin. Relatively fewer lignin but more soluble phenols from original rape straw were removed by LeLac. The enhancement of enzymatic hydrolysis in original rape straw should be a combined result of polyphenols removal and lignin degradation caused by LeLac. This study demonstrated that the LeLac could improve the utilization of rape straw by degrading its lignin, meanwhile it’s worth noting that removing the soluble phenols by LeLac might also play an important role.