黄腐酸(FA)发酵生产及分离提取的研究
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摘要
本文对苹果渣固体发酵生产黄腐酸工艺及产物分离提取条件进行了优化研究。采用正交实验,连续二步固体发酵工艺的实验方法,实验表明发酵的最佳物料比为果渣:豆饼:麸皮=50:4:3,含水量为20%;发酵最佳工艺先接绿色木霉和康氏木霉各2.5%,28℃,pH=6.2发酵12天,灭菌,再接种假丝酵母、酿造酵母、枯草杆菌和棒杆菌各2.5%,32℃,继续发酵12天,可使黄腐酸产率为41.2%。
     在对发酵苹果渣中黄腐酸分离提取进行的研究中,先采用碱溶酸析法通过正交实验得出了从发酵产物中提取黄腐酸的最佳条件:WNaOH=3.0%,60℃,萃取1.5h,然后以盐酸调pH=2.0,酸析得到黄腐酸,纯度可达45.8%。另外,利用KMnO4氧化降解法从碱提取液中进一步回收黄腐酸,其条件为:0.25mol/1 KMnO_4用量为KMnO_4溶液:腐植酸钠溶液=1:6.25(体积比),pH=12,85℃,氧化0.5h,可回收到产量1.12g/1的黄腐酸。所以,苹果渣固体发酵制取黄腐酸是一条可行的,经济的,快速的途径。
The technology of solid state fermentation and extraction of Fulvic acid with apple pomace has been studied. The optimized material ratio of apple pomace to bean cake to bran was 50:4:3 containing water up to 20% by the orthogonal experiments and two-step solid state fermentation was used for Fulvic acid production. First, Trichoderma viride and Trichoderma Koningii were inoculated respectively in the ratio of 2.5% in the meantime. The solid substrates were fermented in pH=6.2 at 28 ℃ for 12d and then exterminated. Secondly, Candida spp, Saccharo spp, Bacillus subtilis and Corynebacterium spp were inoculated respectively in the ratio of 2.5% at the same time. The solid substrates were incubated at 32℃ for 12d. The yield of fulvic acid reached to 41.2% under the optimized conditions.
    The orthogonal experiment showed that the optimum conditions of extraction from fermenting products were that density of sodium hydroxide concentration is 3.0%, extracted at 60℃ for 1.5h, then adjusted acidity with hydrochloric acid to pH=2.0, the purity of Fulvic acid was 45.8%. Fulvic acid was further extracted from alkaline extract with potassium permanganate solution in order to oxidate degradation under optimized conditions of the ratio of 0.25mol/l potassium permanganate solution to sodium humate solution by 1:6.25 (voluminal ratio), pH=12, at 85℃,oxidative degradation for 0.5h, the yield of Fulvie acid was 1.12g/l.
    Solid state fermentation of Fulvic acid with apple pomace as the main raw material was fast, doable and economical method.
引文
[1] Hendfickson J B, Gran D T, Hammod G S. Organic Chemistry [M]. Boston: Willard Grand Press 1970, 287-288.
    [2] 郑平.煤炭腐植酸的生产和应用[M].北京:化学工业出版社,1991,52-57.
    [3] Busnot A, Busnot F, LeQuerlerJ F, et al. Characterization of humic substances extracted from different sediments of the lower Normand region [J]. Thermo chimica Acta, 1995,254:319-330.
    [4] 张振民.腐植酸类物质的胶体化学性质[J].江西腐植酸,1981,1:3~11.
    [5] Saiz-Jimenez C. The Chemical Structure of humic substances: recent advances [M]. In: Piccolo A(Ed), Humie Substances in Terrestrial Ecosystems. Elsevier, Amsterdam. 1996, 1~44.
    [6] 姜爱莉.发酵制备黄腐酸的研究[J].烟台大学学报,2000,13(3):205~209.
    [7] Martin J P, Haiderk. A Comparison of the use of phenolase and peroxidase for the synthesis of model humic acid-type polymers [J]. soil science Society of America Journal, 1980, 44: 983~988.
    [8] 叶美德,李维亮.江南麦区应用黄腐酸防病增产的效果试验[J].腐植酸,1991,2:23~24.
    [9] Goh K M, Stevenson F J. Comparison of infrared spectra of Synthetic and natural humic and Fulvic acids [J]. Soil science, 1971, 112: 392~400.
    [10] 杨慧生.北京腐植酸的研究和应用.北京市腐植酸综合利用办公室,1982,129~130.
    [11] Malcolm R E, Vaughan D. Comparative effect of soil organic matter fractions on phosphatase activities in wheat roots [J]. Plant & soil, 1979, 51(1): 117-126.
    [12] 张辉,姜文勇,刘波.不同来源腐植酸促进植物生长活性及作用机理研究.Ⅰ.腐植酸样品制备,结构测定及对植物种子萌发的影响。
    [13] 白燕等.几种常用农药与黄腐酸的急性联合毒性试验[J].腐植酸,1988,(4)26~27.
    [14] 李绪行.黄腐酸增强小麦抗早能力和生理机制初探[J].植物学报,1992,9(2):
    
    44~46.
    [15]程扶玖,杨道麟.腐植酸对小麦抗旱的生理效应[J].应用生理学报,1995,6(4):363~367.
    [16]吉甘生,朱遐玲.抑制蒸腾剂的节水机理及应用技术的研究验收评价报告[J].腐植酸,1996,4:18~30.
    [17]杨晓玲.黄腐酸及小麦专用黄腐酸浸种对小麦育苗生长及干旱条件生理影响[J].腐植酸,1996,3:20~21.
    [18]马瑞昆,贾秀玲.FA对小麦生理作用及在节水栽培上应用[J].腐植酸,1995,3:17~25.
    [19]朱京涛,高宝珍,朱通顺.关于黄腐酸在农业上的四大作用及其相关问题的研讨[J].腐植酸,1997,4:1~4.
    [20]李维琴,王淑英.黄腐酸对防止黄瓜霜霉病增效作用的研究[J].腐植酸,1991,3:27~32.
    [21]李淑捷等.腐植酸结构与植物刺激作用[M].腐植酸化学学术讨论会论文集,1979,170-179.
    [22]郑平等编,黄腐酸类物质在农业与医药中的应用[M],北京:化学工业出版社。
    [23]Mather S P. Microbal utilization of soil humic acid[J]. Plant and soil, 1967, 27:298 ~299.
    [24]王一鸣.黄腐酸——简易有效抗旱剂[J].腐植酸,1991,1:32~34.
    [25]李卓杰,刘琼等.腐植酸对植物生长的影响[J].腐植酸,1991,1:38~39.
    [26]王天立.黄腐酸在农业上的应用[J].腐植酸,1992,1:14~16.
    [27]许旭旦等.叶面喷施抗早剂一号提高小麦抗旱能力及其增产效果的研究[J].1984,2:53~68.
    [28]王天立等.我国在腐植酸类物质抗蒸腾作用方面的研究进展[J].腐植酸,1996,3:1-7.
    [29]陈荣峰等.葡萄多效增糖灵(FiG)的研制及其应用效果的研究[J].江西腐植酸,1986,1:26~35.
    [30]黄云祥,朱京涛.绿丰收对黄瓜生长及产量的影响[J].腐植酸,1997,1:23-28.
    
    
    [31]何立千主编.生物技术黄腐酸的研究和应用[M].北京:化学工业出版社,1999,218~226.
    [32]黄云,邹宗杰.腐植酸对油白菜产量,品质效应的研究[J].腐植酸,1994,1:23~24.
    [33]姚发业等.黄腐酸抗炎作用的研究[J].腐植酸,1988,2:42~43.
    [34]陈祥银等.黄腐酸钠对大鼠角叉胶致胸膜炎的作用研究[J].生理科学,1988,8(1):37-39.
    [35]唐国藩等.黄腐酸钠治疗各种角膜溃疡32例研究[J].腐植酸,1989,4:42-43.
    [36]王德民等.北京医学院学报,1985,17(1):15-17.
    [37]李绍兰等.吉林石油化工,1982,(3):47-49.
    [38]赵和等.中国酿造,1996,(2):26-28.
    [39]秦万德.腐植酸的综合利用[M].北京:科学出版社,1987,226-228.
    [40]张德和,杨国仪,朱恩兰等.用沸点法测定黄腐酸的分子量[J].化学通报,1981,11:17-20.
    [41]尹淑芝.碱溶、离子交换树脂混合法提取黄腐酸[J].化学工程师,1994,45(6):53-54.
    [42]裘余丹.煤中黄腐酸的电渗析法分离纯化工艺方法研究[J].腐植酸,1999,4:19-23.
    [43]刘大强,刘桂缓,蒋崇菊.提高黄腐酸产率的实验研究[J].哈尔滨科学技术大学学报,1996,20(1):106-109.
    [44]李全民.发酵肥专利配方[P].中国专利:CN87104531.1987-9-10.
    [45]于建国,孙英昌,孙玉凤.腐植酸营养液及其制备方法[P].中国专利:CN94104441.1993-10-4.
    [46]卢庆萍等.秸杆处理技术的研究[J].中国饲料,1999,10:26-28.
    [47]边文骅,边志立等.利用发酵工艺生产黄腐酸方法[P].中国专利:CN92112965.1994-05-25.
    [48]张常书.生化黄腐酸的界定及其资源的开发利用探讨[J].腐植酸,2000,1:15-19.
    [49]Andereas kappler, Rongji, Andreas Brune. Synthesis and characterization of specifically ~(14)C-Labeled humic model compounds for feeding trial with
    
    soil-feeding termits[J]. Soil Biochemistry, 2000, 32:1271~1273.
    [50] Haider K. The synthesis and degradation of humic substances in soil. Transaction of the 8th Congress of the Intemational Soil Science Society. 1987, 6: 644-656.
    [51] Gomes R C, Mangrieh A S, Coelho R R R. et al. Elemental, functional group and infrared spectroscopic analysis of aetinomycete melanins from Brazilian solis[J]. Biology and Fertility, 1996, 21: 84-88.
    [52] Stevenson F J. Humus Chemistry [M]. Genesis, Composition, Reactions, wiley, NewYork, 1994, 2: 1680-1683.
    [53] Chin Y P, Aiken G, Lough Lin E O. Molecular weight, polydispersity, and spectroscopic properties of aquatic humic substances [J]. Environ sci Technol, 1994, 28: 1853-1858.
    [54] Ladd J N, Butler J H A. Comparison of some properties of soil humic acids and Synthetic phenolic polymers incorpsration amino derivatives[J]. Australian Journal of soil Research, 1996, 4:41-54.