赵国群, 牛梦天, 卢士康, 关军锋. 梨渣固态发酵培养多粘类芽孢杆菌的工艺[J]. 农业工程学报, 2016, 32(7): 303-308. DOI: 10.11975/j.issn.1002-6819.2016.07.043
    引用本文: 赵国群, 牛梦天, 卢士康, 关军锋. 梨渣固态发酵培养多粘类芽孢杆菌的工艺[J]. 农业工程学报, 2016, 32(7): 303-308. DOI: 10.11975/j.issn.1002-6819.2016.07.043
    Zhao Guoqun, Niu Mentian, Lu Shikang, Guan Junfeng. Cultivation of Paenibacillus polymyxa by solid-state fermentation of pear residues[J]. Transactions of the Chinese Society of Agricultural Engineering (Transactions of the CSAE), 2016, 32(7): 303-308. DOI: 10.11975/j.issn.1002-6819.2016.07.043
    Citation: Zhao Guoqun, Niu Mentian, Lu Shikang, Guan Junfeng. Cultivation of Paenibacillus polymyxa by solid-state fermentation of pear residues[J]. Transactions of the Chinese Society of Agricultural Engineering (Transactions of the CSAE), 2016, 32(7): 303-308. DOI: 10.11975/j.issn.1002-6819.2016.07.043

    梨渣固态发酵培养多粘类芽孢杆菌的工艺

    Cultivation of Paenibacillus polymyxa by solid-state fermentation of pear residues

    • 摘要: 为了充分利用梨渣这一资源,该文以新鲜鸭梨渣为主要原料,采用固态发酵方式,研究了梨渣与麸皮比例、外加氮源、无机盐、装瓶量、培养温度和时间对生防用多粘类芽孢杆菌D1生长及产芽孢的影响。研究发现,梨渣是一个良好的固态发酵原料,适合培养多粘类芽孢杆菌。当梨渣与麸皮的质量比为5∶2时,获得最大活细胞数,为2.5×109 cfu/g。向梨渣发酵培养基中添加尿素、硝酸钠和豆饼粉可明显提高活菌数和芽孢数,其中添加豆饼粉的效果较好。培养基中添加质量分数为0.5%磷酸二氢钾极大地促进了菌体生长和芽孢的形成。随着培养基装瓶量的增加,芽孢数也随之明显减少。多粘类芽孢杆菌D1的较适培养温度为34°C,较适培养时间为120 h。适宜培养条件下的活菌数、芽孢数和芽孢率分别为5.09×109 cfu/g、4.78×109 cfu/g和93.9%。该研究结果为利用梨渣发酵生产多粘类芽孢杆菌菌剂提供参考。

       

      Abstract: Abstract: Pear juice is one of main processed products of pear fruit in China. A lot of pear residues are produced during pear juice processing, and the waste is up to 40%-50% of pear fruit weight. Because pear residues contain many stone cells and can not be used directly as animal feed, most of them have to be discarded in fields or near factories. Paenibacillus polymyxa is a widely distributed bacterial species in the environment, and has no pathogen to humans, animals and plants. P. polymyxa strain produces several antibiotics and hydrolytic enzymes, which play an important role in the biocontrol of plant pathogens and the promotion of plant growth. It is reported to be both a plant-growth-promoting and effective biocontrol agent. P. polymyxa D1, a naturally occurring biocontrol agent isolated from soil, can stimulate plant growth and effectively suppresse several soil-borne diseases. Many researches on submerged fermentations of P. polymyxa have been done, but study on its solid-state fermentation has not been reported at the present. In this work, fresh Yali pear’s residues were used to cultivate P. polymyxa D1 in a solid-state fermentaion. Fresh Yali pear’s residues contained 76.2% moisture, 5.5% residual sugars and 0.32% proteins. The effects of cultivation conditions on its cell growth and sporulation were investigated. The results showed that pear residues were good material for solid-state fermentation and very suitable to cultivate P. polymyxa D1. The mositure of fresh Yali pear’s residues was very high, which might not be in favour of the cell growth during solid-state fermentation. Addition of wheat bran into pear residues improved significantly the cell growth. The possible reason of this result was that oxygen supply and nutritional status of the culture medium were improved obviously when pear residues were mixed with wheat bran. When the ratio between pear residue and wheat bran was 5:2, the highest viable cell number was achieved, 2.5×109 cfu/g. Spore number was an important parameter of microbial agent. Cultivation conditions could influence bacterial sporulation. The viable cell number and spore yield of P. polymyxa D1 obviously increased when urea, sodium nitrate and soybean meal were added into the cultural medium of pear residues, and soybean meal had the best stimulation effect. However, NH4NO3, NH4Cl and degossypolized cottonseed protein inhibited the cell growth and spore formation. Mineral salts also influenced the cell growth and sporulation of P. polymyxa D1. Supplement of NaCl, MgSO4, CaCO3 and FeSO4 into the medium largely inhibited the spore formation of P. polymyxa D1. Addition of KH2PO4 greatly promoted the cell growth and sporulation, and its spore yield and sporulation efficiency were 4.52×109 cfu/g and 95.6%, respectively. P. polymyxa is a highly anaerobic bacterium. The spore yield of P. polymyxa D1 decreased obviously as the medium volume in the fermenter increased. The optimal cultivation temperature of P. polymyxa D1 was 34°C, which was higher than that in submerged frementation. The optimal cultivation time of P. polymyxa D1 was 120 h. It was found that the spores were formed very slowly by the cells of P. polymyxa D1 when cultivated with pear residues in solid-state fermentaion, compared with submerged fermentation.

       

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