菌/酶添加对甜高粱渣青贮预处理作用的强化效果

    Improvement for the ensiling pretreatment effectiveness of sweet sorghum bagasse by fortified with Lactobacillus plantarum and cellulolytic enzymes

    • 摘要: 为实现生物质甜高粱渣的青贮强化预处理和能源化利用,该研究探究了不同添加剂对其青贮质量和酶解糖化效果的影响。试验设置对照组(CK组)、植物乳杆菌组(L组)、纤维复合酶组(E组)和复合添加剂组(LE组),系统考察甜高粱渣在21 d青贮预处理期间的营养成分、木质纤维组分和发酵品质的动态变化,采用隶属函数法评价青贮质量,利用高通量测序技术分析青贮预处理过程的微生物菌群动态演绎。结合酶解糖化性能评价青贮预处理作用的强化效果,从而筛选适宜的添加剂。结果表明:青贮预处理后甜高粱渣的粗蛋白、淀粉、中性洗涤纤维、酸性洗涤纤维、半纤维素和纤维素等能量组分含量显著高于原料(P<0.05)。青贮21d时,3个添加剂组的干物质、可溶性碳水化合物和粗蛋白含量显著高于CK组(P<0.05),综纤维素含量显著低于CK组(P<0.05),且E组的干物质含量最高,L组的粗蛋白含量最高,E组和LE组的可溶性碳水化合物含量最高。青贮预处理期间,3 个添加剂组的pH值均低于4.2,L组的乳酸和乙酸含量明显高于CK组(P<0.05),E组的乳酸/乙酸和乳酸/总有机酸比值显著高于CK组(P<0.05)。隶属函数法综合评价E组的青贮发酵质量最高。微生物菌群结果显示,甜高粱渣经青贮预处理后,3个添加剂组的细菌菌群丰富度和多样性都明显下降,门水平细菌主要以变形菌为主,属水平细菌主要包含肠杆菌、泛菌、明串珠菌、魏斯氏菌、拉恩氏菌和葡糖杆菌等,这些微生物与青贮质量密切相关。甜高粱渣经青贮预处理后的还原糖得率显著提升,尤其E组得率比原料提高了117%。结合成本效益法分析,利用纤维复合酶对甜高粱渣进行青贮强化处理的E组纯收益最高,较甜高粱渣原料提高了近三倍。总之,单独添加纤维复合酶不仅能明显改善甜高粱渣的青贮发酵质量,实现稳定保存,还能使其青贮预处理作用得到强化,进而提高甜高粱渣的生物降解性能,是一种经济合理、技术可行的青贮预处理强化方法。

       

      Abstract: Abstract: Sweet sorghum bagasse (SSB) is the resultant waste after extraction of sugar-rich juice from the stalks during bioethanol production. As a typical biomass, the SSB consists of moisture, unspent soluble sugar, and abundant lignocellulosic component (cellulose, hemicelluloses and lignin). The undisposed SSB can inevitably cause the environmental pollution and resource waste. Hence, the effective storage of SSB is necessary for the year-round stable operation of biofuel plants. Among them, ensiling (wet storage) can be an efficient technology available for the SSB preservation and utilization, which enable the SSB to be processed all year around. Moreover, ensiling can also act as a pretreatment strategy, due to the benefits for the cell wall degradation and the improvement of biomass bioconversion for subsequent processing. In this study, the potential of ensiling pretreatment was investigated to fortify with Lactobacillus plantarum inoculant and cellulolytic enzymes on the improvement of ensiling quality to modulate the performance of enzymatic saccharification of SSB. The SSB were ensiled with no additives (CK), Lactobacillus plantarum (L), cellulolytic enzyme (E), and a combination of L and E (LE) for 21 days. The dynamic changes of nutrient composition, lignocellulosic components, and ensiling fermentation characteristics were investigated, and then dynamic evolution of the bacterial community structure was analyzed by high-throughput sequencing at HiSeq2500 platform. Ensiling quality was comprehensively evaluated by membership function method, in order to screen the suitable silages additives in consideration of enzymatic hydrolysis performance. The results showed that the addition of Lactobacillus plantarum inoculant and cellulolytic enzymes either alone or in combination was facilitated the lactic acid fermentation to reduce the fermentation losses, as evidenced by the higher content of crude protein, starch, neutral detergent fiber, acid detergent fiber, hemicellulose and cellulose components than that of un-ensiled SSB. The content of dry matter, crude protein and water-soluble carbohydrates in silages with additives were higher than those in the CK, but the content of holocellulose lower. Furthermore, the content of dry matter in the E silages, the content of crude protein in the L silages, and the content of water-soluble carbohydrates in the LE silages were the highest in all silages, respectively. The pH value of all silages significantly decreased to below 4.2, accompanied by the ratio of lactic Acid (LA) and total organic acids (TOA) higher than 0.65 and the ratio of lactic acid (LA) and acetic acid (AA) higher than 2 during the whole ensiling for 21 days. The content of LA and AA in the L silages were significantly higher than those of silages at the CK (P<0.05), the ratio of LA/TOA and LA/AA in the E silages were significantly higher than those of silages at the CK(P<0.05). The membership function analysis indicated that the silages at the E group shared the highest comprehensive scores at 21 days. The bacterial community structure showed that the Proteobacteria was the main bacteria in all silages during ensiling fermentation. At 21 days of ensilage, Enterobacteriaceae, pantoea and leuconostoc were the main species in the silages at genus level. ACE, Chao1 and Shannon index in the E, L and LE silages were significantly lower than those of CK group (P<0.05). It infers that the ensiling pretreatment with the additives was effectively reduced the microbial richness and diversity. Principal Coordinate Analysis (PCA) was constructed using the relative abundance of bacteria at the genus level, indicating the outstanding separation in the SSB before and after ensiling pretreatment. After ensiling pretreatment, the yield of reducing sugar in all SSB silages significantly increased, compared with the un-ensiled SSB. Consequently, the reducing sugar yield of in the E silages increased by 117%. The highest net income of E silages fortified with cellulolytic enzyme was almost three-fold that of un-ensiled SSB using cost-benefit analysis. In conclusion, the ensiling pretreatment fortified by cellulolytic enzymes can be expected to serve as a cost-effective, eco-friendly and tech-feasible strategy for the preservation and pretreatment of SSB biomass. Especially, cellulolytic enzyme can be used to modulate the ensiling pretreatment performance. Therefore, the ensiling quality and the biodegradation performance of SSB silage can be effectively improved for the bioenergy utilization of sweet sorghum bagasse.

       

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