• 中文核心期刊
  • CSCD来源期刊
  • 中国科技核心期刊
  • CA、CABI、ZR收录期刊

纤维素降解菌产酶条件优化及其堆肥效果研究

Enzyme Generation and Effect in Composting of Cellulolytic Bacteria

  • 摘要:
    目的 筛选出适用于青海地区纤维素类物质降解的菌株,提高堆肥效果。
    方法 以青海省海北州森林土壤为样品,采用刚果红染色法和3,5-二硝基水杨酸(3,5-Dinitrosalicylic acid, DNS)法筛选高效纤维素降解菌株,利用生理生化试验和16S rDNA序列分析对菌株进行鉴定,并优化其发酵产纤维素酶条件,制成菌剂后按1%的接种量接种于堆肥试验中,测定堆肥各项腐熟指标。
    结果 筛选出的高活性降解菌MX-1-1为蕈状芽胞杆菌(Bacillus mycoides),该菌株羧甲基纤维素酶、滤纸酶和外切-β-葡聚糖酶活性分别为24.95、11.36、16.15 U·mL−1。最适产羧甲基纤维素酶发酵条件:氮源牛肉膏、碳源CMC-Na、培养基初始pH 5.0、发酵温度20 ℃、培养时间3 d,优化后羧甲基纤维素酶活力提高了51.5%。堆肥过程中,添加MX-1-1菌剂的堆体升温快,高温期持续时间长,pH波动幅度小。堆肥结束时,堆体电导率为0.96 ms·cm−1,含水量最低,硝态氮和铵态氮含量均达腐熟标准,全氮和有机质含量优于未添加菌剂堆体和D50菌剂堆体。
    结论 菌株MX-1-1具有较强的纤维素降解能力,能够促进堆体微生物代谢,加快堆体腐熟,并提高堆肥品质。

     

    Abstract:
    Objective Microbes capable of degrading cellulose in compost and suitable for application in Qinghai-Tibet Plateau region were tested, and fermentation conditions to maximize the enzyme generation optimized.
    Method Soil samples were collected at the forests in Haibei Prefecture, Qinghai Province. Cellulolytic bacteria were isolated by the Congo red staining and 3,5- Dinitrosalicylic acid (DNS) methods. Highly efficient cellulose-degrading strains were identified by physiochemical tests and 16S rDNA sequencing. Conditions to effectively generate cellulase by the bacteria were optimized. Extracts of the enzymes produced from the fermentation were diluted to 1% and added to compost with cellulolytic process and compositing conditions monitored.
    Result The isolated MX-1-1, which was subsequently identified as a strain of Bacillus mycoides, displayed the greatest degradation activities on carboxymethyl cellulose, filter paper, and exo-β-glucanose at 24.95, 11.36, and 16.15 U·mL−1, respectively. In generating carboxymethyl cellulase, the fermentation that increased 51.5% in activity over control employed beef paste for nitrogen and CMC-Na for carbon in a medium of an initial pH of 5.0 at 20 ℃ for 3 d. With the addition of MX-1-1 extract, the compost rose rapidly to a lasting high temperature with relatively stable pHs. At the end of fermentation, the compost showed a conductivity of 0.96 ms·cm−1 with the moisture content at its lowest level, the nitrate and ammonium nitrogen at the matured decomposition level, and the total nitrogen and organic matter higher than those in asepsis or D50 compost.
    Conclusion MX-1-1, a strain of Bacillus mycoides, was found highly cellulolytic. By adding it for composting farm waste, the metabolic activity was significantly enhanced to hasten the cellulose decomposition.

     

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