Wei Ya-Qin, Long Rui-Jun, Yang Hui, Yang Hong-Jian, Shen Xi-Hui, Shi Rui-Fang, Wang Zhi-Ye, Du Jun-Guo, Qi Xiao-Jin, Ye Qian-Hong
Institute of Biological Research, Gansu Academy of Sciences, Lanzhou, 730000, PR China; School of Life Sciences, Lanzhou University, Lanzhou, 730000, PR China.
School of Life Sciences, Lanzhou University, Lanzhou, 730000, PR China.
Anaerobe. 2016 Jun;39:158-64. doi: 10.1016/j.anaerobe.2016.03.005. Epub 2016 Mar 12.
Several natural anaerobic fungus-methanogen co-cultures have been isolated from rumen and feces source of herbivores with strong fiber degrading ability. In this study, we isolated 7 Neocallimastix with methanogen co-cultures from the rumen of yaks grazing on the Qinghai Tibetan Plateau. Based on morphological characteristics and internal transcribed spacer 1 sequences (ITS1), all the fungi were identified as Neocallimastix frontalis. The co-cultures were confirmed as the one fungus - one methanogen pattern by the PCR-denatured gradient gel electrophoresis (DGGE) assay. All the methanogens were identified as Methanobrevibacter ruminantium by 16s rRNA gene sequencing. We investigated the biodegrading capacity of the co-culture (N. frontalis + M. ruminantium) Yaktz1 on wheat straw, corn stalk and rice straw in a 7 days-incubation. The in vitro dry matter digestibility (IVDMD), acid detergent fiber digestibility (ADFD) and neural detergent fiber digestibility (NDFD) values of the substrates in the co-culture were significantly higher than those in the mono-culture N. frontalis Yaktz1. The co-culture exhibited high polysaccharide hydrolase (xylanase and FPase) and esterase activities. The xylanase in the co-culture reached the highest activity of 12500 mU/ml on wheat straw at the day 3 of the incubation. At the end of the incubation, 3.00 mmol-3.29 mmol/g dry matter of methane were produced by the co-culture. The co-culture also produced high level of acetate (40.00 mM-45.98 mM) as the end-product during the biodegradation. Interestingly, the N. frontalis Yaktz1 mono-culture produced large amount of lactate (8.27 mM-11.60 mM) and ethanol (163.11 mM-242.14 mM), many times more than those recorded in the previously reported anaerobic fungi. Our data suggests that the (N. frontalis + M. ruminantium) Yaktz1 co-culture and the N. frontalis Yaktz1 mono-culture both have great potentials for different industrial use.
已从具有强大纤维降解能力的食草动物瘤胃和粪便来源中分离出几种天然厌氧真菌-产甲烷菌共培养物。在本研究中,我们从青藏高原放牧的牦牛瘤胃中分离出7种与产甲烷菌共培养的新美鞭菌属真菌。根据形态特征和内部转录间隔区1序列(ITS1),所有真菌均被鉴定为额叶新美鞭菌。通过PCR-变性梯度凝胶电泳(DGGE)分析,确认共培养物为一种真菌-一种产甲烷菌模式。通过16s rRNA基因测序,所有产甲烷菌均被鉴定为反刍短杆菌。我们研究了共培养物(额叶新美鞭菌+反刍短杆菌)Yaktz1在7天培养期内对小麦秸秆、玉米秸秆和稻草的生物降解能力。共培养物中底物的体外干物质消化率(IVDMD)、酸性洗涤纤维消化率(ADFD)和中性洗涤纤维消化率(NDFD)值显著高于单培养的额叶新美鞭菌Yaktz1。共培养物表现出高多糖水解酶(木聚糖酶和FPase)和酯酶活性。共培养物中的木聚糖酶在培养第3天对小麦秸秆的活性达到最高,为12500 mU/ml。在培养结束时,共培养物产生了3.00 mmol-3.29 mmol/g干物质的甲烷。共培养物在生物降解过程中还产生了高水平的乙酸盐(40.00 mM-45.98 mM)作为终产物。有趣的是,单培养的额叶新美鞭菌Yaktz1产生了大量的乳酸(8.27 mM-11.60 mM)和乙醇(163.11 mM-242.14 mM),比先前报道的厌氧真菌中的记录高出许多倍。我们的数据表明,(额叶新美鞭菌+反刍短杆菌)Yaktz1共培养物和额叶新美鞭菌Yaktz1单培养物在不同的工业用途中都具有巨大潜力。