农业固体废物堆肥中木质素的微生物降解研究
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摘要
利用微生物对农业废物进行堆肥化是当前处理农业废物的一大趋势。堆肥过程中微生物的种类、数量及群落结构是决定该生态体系的物质循环和废物特别是木质素类物质降解能力的重要因素。由于环境微生物的可培养性差,传统的培养技术不适用堆肥微生物群体组成和结构的解析。近年来发展起来的醌指纹法,利用微生物体内广泛存在的类异戊二烯醌作为标记物分析微生物群落,成为环境微生物和微生物生态领域的研究热点。本文详细介绍了醌指纹法整个体系的建立过程,并利用该体系对堆肥过程中的微生物群落及具有木质素降解潜能的微生物群落进行了成功分析。在堆肥过程中MK7(H4)、Q7、Q8含量较多,表明γ-变形菌和放线菌在整个过程中起主要作用,同时堆肥化不同时期的群落变化基本符合传统的研究结果。而在具有木质素降解潜能的微生物群落中,主要醌种为MK7(H2)、MK7(H4)、Q9(H2),表明堆肥化过程中降解木质素的主要是一些真菌和γ-变形菌。降解主要发生在高温期和二次发酵时期。微生物群落多样性指数较小,说明物种都比较单一,同时群落结构随着堆制时间发生巨大变化。
本文还研究了生物表面活性剂鼠李糖脂对黄孢原毛平革菌、简青霉产木质素降解酶能力的影响。结果表明,鼠李糖脂对酶活性的影响与鼠李糖脂的浓度、投加形式、菌种及酶的种类有关。0.0065%的鼠李糖脂不同程度地促进了黄孢原毛平革菌产LiP酶,MnP酶,简青霉产LiP酶,漆酶,0.013%的鼠李糖脂可以促进黄孢原毛平革菌产LiP酶,简青霉产漆酶,但却明显抑制了另外两种酶的活性;鼠李糖脂可以使简青霉产MnP的高峰期提前,但不能提高酶活性。通过影响产酶能力,鼠李糖脂使两株菌对木质素的降解率均高于对照样。此外,接种鼠李糖脂产生菌——铜绿假单胞菌的效果达到甚至超过添加鼠李糖脂的效果。
It is a current trend to degrade agriculture waste in composting process using different microorganism. The species (composting), biomass (richness) and the community (structure) of microorganism are dominating factors in composting. The traditional cultivating method is falling into disuse since its high deviation. However, the quinine profile analysis can overcome which based on the different kind of isoprenoid quinine in microorganism. This paper illuminated the establishment of the quinine analysis system and successfully using it to characterize the microbial community in compost. The microbial community having the potential to degrade lignin was also investgated. The major quinine species in compost process were MK-7(H4), Q7 and Q8, indicating thatγ-subclass Proteobacteria and actinomycetes are the major microorganism. The results of community changing in different compost period that obtained through the quinine ananlysis accorded with that obtained through traditional cultivation. In the community having the potential of degrading lignin, MK-7(H2), MK-7(H4) and Q9(H2) were found to be the major quinines, which mostly represent some fungi and Proteobacteria. The microbes worked mostly in the high-temperature period and the second fermenting period. Both of the two microbial communities had relatively small diversity of quinine (DQ), which indicated the singleness of the microbial species.
The effects of biosurfactant rhamnolipid on the enzyme production by Phanerochaete chrysosporium and Penicillium simplicissimum were also studied. Results show that the effects were connected with the concentration of rhamnolipid, adding ways, fungi species and type of enzymes. Rhamnolipid at 0.0065% increased the activities of LiP, MnP by P. chrysosporium, LiP, laccase by P. simplicissimum, while rhamnolipid at 0.013% only stimulated the production of LiP by P. chrysosporium, laccase by P. simplicissimum, but restrained the other two enzymes’activities. However, rhamnolipid only made the highest MnP of P. simplicissimum coming ahead of which of the control, with no change of MnP activity. All the degradation rate of lignin which with rhamnolipid was higher than that of the control, and inoculation of Pseudomonas aeruginosa showed the same or even better stimulative effect comparing with those rhamnolipid was added.
本文还研究了生物表面活性剂鼠李糖脂对黄孢原毛平革菌、简青霉产木质素降解酶能力的影响。结果表明,鼠李糖脂对酶活性的影响与鼠李糖脂的浓度、投加形式、菌种及酶的种类有关。0.0065%的鼠李糖脂不同程度地促进了黄孢原毛平革菌产LiP酶,MnP酶,简青霉产LiP酶,漆酶,0.013%的鼠李糖脂可以促进黄孢原毛平革菌产LiP酶,简青霉产漆酶,但却明显抑制了另外两种酶的活性;鼠李糖脂可以使简青霉产MnP的高峰期提前,但不能提高酶活性。通过影响产酶能力,鼠李糖脂使两株菌对木质素的降解率均高于对照样。此外,接种鼠李糖脂产生菌——铜绿假单胞菌的效果达到甚至超过添加鼠李糖脂的效果。
It is a current trend to degrade agriculture waste in composting process using different microorganism. The species (composting), biomass (richness) and the community (structure) of microorganism are dominating factors in composting. The traditional cultivating method is falling into disuse since its high deviation. However, the quinine profile analysis can overcome which based on the different kind of isoprenoid quinine in microorganism. This paper illuminated the establishment of the quinine analysis system and successfully using it to characterize the microbial community in compost. The microbial community having the potential to degrade lignin was also investgated. The major quinine species in compost process were MK-7(H4), Q7 and Q8, indicating thatγ-subclass Proteobacteria and actinomycetes are the major microorganism. The results of community changing in different compost period that obtained through the quinine ananlysis accorded with that obtained through traditional cultivation. In the community having the potential of degrading lignin, MK-7(H2), MK-7(H4) and Q9(H2) were found to be the major quinines, which mostly represent some fungi and Proteobacteria. The microbes worked mostly in the high-temperature period and the second fermenting period. Both of the two microbial communities had relatively small diversity of quinine (DQ), which indicated the singleness of the microbial species.
The effects of biosurfactant rhamnolipid on the enzyme production by Phanerochaete chrysosporium and Penicillium simplicissimum were also studied. Results show that the effects were connected with the concentration of rhamnolipid, adding ways, fungi species and type of enzymes. Rhamnolipid at 0.0065% increased the activities of LiP, MnP by P. chrysosporium, LiP, laccase by P. simplicissimum, while rhamnolipid at 0.013% only stimulated the production of LiP by P. chrysosporium, laccase by P. simplicissimum, but restrained the other two enzymes’activities. However, rhamnolipid only made the highest MnP of P. simplicissimum coming ahead of which of the control, with no change of MnP activity. All the degradation rate of lignin which with rhamnolipid was higher than that of the control, and inoculation of Pseudomonas aeruginosa showed the same or even better stimulative effect comparing with those rhamnolipid was added.
引文
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