Isolation and Composting Efficiency of Cellulose-degrading Fungus from Litter Fermentation Bed in Pig Pan
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摘要: 采用CMC-Na平板法和刚果红染色法从猪发酵床陈化垫料中分离到1株纤维素高效降解真菌M6。该菌株测序后的ITS基因序列与NCBI数据库进行BLAST比对，并构建系统发育树，该菌株初步鉴定为草酸青霉。优化固态发酵产酶工艺，确定最佳产酶条件：以麸皮+羧甲基纤维素钠为碳源，豆粕为氮源，初始pH值为6.0，接种量为10%，发酵温度为30℃、发酵时间为96 h，羧甲基纤维素酶酶活最高可达1 446 U·g-1，是优化前的2.31倍。堆肥试验结果表明：接种M6组和EM菌剂组均在第3 d进入高温期（>50℃），且高温分别维持了10 d和6 d；菌株M6组、EM组、对照组纤维素降解率分别为41.1%、38.8%、14.8%。因此，菌株M6组高温维持时间长、纤维素降解率高，在发酵床陈化垫料堆肥腐熟发酵中具有潜在的应用前景。Abstract: A fungus, M6, that displayed a powerful ability to degrade cellulose was isolated from the waste fermentation beds at pig farms using the CMC-Na plate and Congo red staining methods. ITS sequence of the isolated strain was blasted with those from NCBI database, and the phylogenetic tree constructed to arrive at a preliminary identification for the fungus as Penicillium oxalate. Subsequently, an optimized enzyme-producing fermentation process on a solid medium inoculated with the isolated fungi was established. It included the application of bran and CMC-Na as the carbon source, soybean meal as the nitrogen source, pH 6.0 at the start, 10% inoculation rate, and fermentation at 30℃ for 96 h. The carboxymethyl cellulase activity of the material increased 2.31 times due to the optimization that reached 1 446 U·g-1. In a composting experiment on the fermentation beds inoculated with M6, EM or control, M6 and EM raised the bed temperature to greater than 50℃ on the 3rd day after start of fermentation, and maintained at the level for 10 d and 6 d, respectively. The cellulose degradation rate was 41.1% with M6, 38.8% with EM, and 14.8% with control. It suggested that, among the 3 tested fungi, M6 was most efficient on the waste treatment for pig farms.
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图 4 不同碳源对菌株M6产酶效果影响
注：图中碳源A为麸皮，B为稻壳粉，C为羧甲基纤维素钠，D为微晶纤维素，E为淀粉，F为葡萄糖，G为蔗糖，H为麸皮+羧甲基纤维素钠。
Figure 4. Effects on enzyme production of M6 cultured with different carbon sources
图 5 不同氮源对菌株M6产酶效果影响
Figure 5. Effects on enzyme production of M6 cultured with different nitrogen sources
图 6 不同培养时间对菌株M6产酶影响
Figure 6. Effects on enzyme production of M6 under different culture 13:09:39
表 1 不同起始pH对菌株M6产酶影响
Table 1. Effects on enzyme production of M6 cultured with different initial pH
pH 3 4 5 6 7 8 9 CMCase酶活力/(U·g-1) 243 581 627 720 387 124 112 
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表 2 不同培养温度对菌株M6产酶影响
Table 2. Effects on enzyme production of M6 cultured under varied temperatures
温度/℃ 20 25 28 30 35 40 45 50 CMCase酶活力/(U·g-1) 157 413 627 846 541 378 176 146
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表 3 不同接种量对菌株M6产酶影响
Table 3. Effects on enzyme production of M6 cultured with different inoculum concentration
不同接种量/% 2 4 8 10 12 14 CMCase酶活力/(U·g-1) 210 374 598 635 432 127
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