高效降解棉籽粕中游离棉酚菌株的筛选及复配发酵方式的优化
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摘要
利用酵母菌和乳酸菌发酵棉籽粕,研究其对棉籽粕中游离棉酚降解率的影响,并对发酵前后棉籽粕的营养指标如活菌数、酸溶蛋白、pH和游离棉酚含量等进行比较。筛选出了可高效降解游离棉酚的酵母菌BLCC4-0327,可改善发酵风味的乳酸菌BLCC2-0092。并对筛选出的酵母菌和乳酸菌复配发酵进行了初步研究,结果表明,最优复配发酵组为BLCC4-0327+BLCC2-0092(1:1),30℃厌氧发酵。与空白对照组相比,最优复配发酵组各发酵阶段的发酵棉籽粕的pH均显著降低(P<0. 05),发酵24 h时pH降至5. 73;酸溶蛋白含量显著提高(P <0.05),发酵24 h时酸溶蛋白质量分数达到14.61%,发酵48 h时酸溶蛋白质量分数达到17.61%;游离棉酚含量显著降低(P<0. 05),发酵24 h时游离棉酚降解率达到70. 99%,发酵48 h时游离棉酚降解率达到73. 44%。由此可知,酵母菌BLCC4-0327与乳酸菌BLCC2-0092复配发酵可有效降低发酵棉籽粕中的游离棉酚含量并改善其营养品质。
This experiment was conducted for studying the effects of cottonseed meal fermented by Lactobacillus and Yeast on free gossypol-degrading rate of cottonseed meal, and comparing the nutritive quality indices of cottonseed meal before and after fermentation, such as viable count of bacteria, pH, acid-soluble protein content and gossypol content, etc. The results showed that the Yeast BLCC4-0327 had high efficient free gossypol-degrading capacity and the Lactobacillus BLCC2-0092 had improving fermentation flavor capacity. And then the mixed fermentation of Lactobacillus and Yeast were studied, the results showed that the optimal fermentation of Yeast BLCC4-0327 and Lactobacillus BLCC2-0092 was 1: 1, 37℃ for aerobic fermentation. Compared with control check group, the pH of fermented cottonseed meal in each fermentation stage of optimal compound fermentation group was signification decreased(P < 0. 05), and the pH after anaerobic fermentation 24 h reduced to 5. 73; the acid-soluble protein content was significantly increased(P < 0. 05), and the acid-soluble protein content after anaerobic fermentation24 h increased to 14. 61% and 48 h increased to 17. 61%; the feed gossypol content was significantly decreased(P <0. 05), and the gossypol degrading rate after anaerobic fermentation 24 h reached to 70. 99% and 73. 44% for48 h. It could thus be seen that compound fermentation Yeast BLCC4-0327 and Lactobacillus BLCC2-0092 can effectively degrade the content of free gossypol in cottonseed meal and improve the nutritive quality of cottonseed meal.
This experiment was conducted for studying the effects of cottonseed meal fermented by Lactobacillus and Yeast on free gossypol-degrading rate of cottonseed meal, and comparing the nutritive quality indices of cottonseed meal before and after fermentation, such as viable count of bacteria, pH, acid-soluble protein content and gossypol content, etc. The results showed that the Yeast BLCC4-0327 had high efficient free gossypol-degrading capacity and the Lactobacillus BLCC2-0092 had improving fermentation flavor capacity. And then the mixed fermentation of Lactobacillus and Yeast were studied, the results showed that the optimal fermentation of Yeast BLCC4-0327 and Lactobacillus BLCC2-0092 was 1: 1, 37℃ for aerobic fermentation. Compared with control check group, the pH of fermented cottonseed meal in each fermentation stage of optimal compound fermentation group was signification decreased(P < 0. 05), and the pH after anaerobic fermentation 24 h reduced to 5. 73; the acid-soluble protein content was significantly increased(P < 0. 05), and the acid-soluble protein content after anaerobic fermentation24 h increased to 14. 61% and 48 h increased to 17. 61%; the feed gossypol content was significantly decreased(P <0. 05), and the gossypol degrading rate after anaerobic fermentation 24 h reached to 70. 99% and 73. 44% for48 h. It could thus be seen that compound fermentation Yeast BLCC4-0327 and Lactobacillus BLCC2-0092 can effectively degrade the content of free gossypol in cottonseed meal and improve the nutritive quality of cottonseed meal.
引文
[1]周媛媛,朱泉,陈生琴,等.瘤胃微生物发酵棉粕对肉鸭生产性能的影响[J].中国油脂,2016,41(7):24-28ZHOU Y Y,ZHU Q,CHEN S Q,et al. Impact of cottonseed meal fermented by rumen microorganisms on production performance of meat ducks[J]. China Oils and Fats, 2016,41(7):24-28
[2]亓秀晔,谢全喜,陈振,等.高效降解游离棉酚并改善棉籽粕营养品质的菌株筛选[J].动物营养学报,2017,29(9):3258-3266QI X Y,XIE Q X,CHEN Z,et al. Screening of high efficient free gossypol-degrading and improving cottonseed meal nutritive quality strains[J]. Chinese Journal of Animal Nutrition,2017,29(9):3258-3266
[3]陈生琴,陈丽娟,程茂基.高效降解棉酚菌株的选育及发酵棉粕最佳工艺的研究[J].中国油脂,2016.41(4):19-24CHEN S Q,CHEN L J,CHENG M J,et al. Screening of high efficient gossypol-degrading strain andoptimization of cottonseed meal[J]. China Oils and Fats,2016,41(4):19-24
[4]LORDELO M M, CALHOUN M C, DALE N M, et al. Relative toxicity of gossypol enantiomers in laying and broiler breeder hens[J]. Poultry Science,2007,86:582-590
[5]吕云峰,王修启,赵青余,等.棉酚在饲料中安全限量及畜产品中残留研究进展[J].中国农学通报,2010,26(24):1-5LYU Y F,WANG X Q,ZHAO Q Y,et al. Research situation on gossypol safety limit in feed and gossypol residues in livestock product[J]. Chinese Agricultural Science Bulletin,2010,26(24):1-5
[6]YANG X, SUN J Y, GUO J L, et al. Identification and proteomic analysis of a novel gossypol-degrading fungal strain[J]. Journal of the Science of Food Agriculture, 2012,92(4):943-951
[7]SUN H, TANG J W, YAO X H, et al. Improvement of the nutritional quality of cottonseed meal by Bacillus subtilis and the addition of papain[J]. International Journal of Agriculture and Biology, 2012, 14(4):563-568
[8]DAI C H,ZHANG L P,MA H L. Ultrasound-assisted detoxification of free gossypol from cottonseed meal[J]. Journal of Food Process Engineering, 2015. 7,https://doi.org/10. 1111/jfpe. 12265
[9]张煜,石常友,王成,等.菌酶协同发酵改善玉米-豆粕型饲料营养价值的研究[J].中国粮油学报,2018,33(3):70-77ZHANG Y,SHI C Y,WANG C,et al. Improving nutrition value of maize-soybean meal based diet by microbial fermenta-tion combined with multiple-enzymes[J]. Journal of the Chinese Cereals and Oils Association,2018,33(3):70-77
[10]张连慧,熊小辉,惠菊,等.发酵豆粕及其在动物养殖行业中的应用研究进展[J].中国油脂,2017,42(3):108-112ZHANG L H,XIONG X H,HUI J,et al. Progress in fermented soybean meal and application in animal breeding industry[J]. China Oils and Fats, 2017,42(3):108-112
[11]张秀林,魏小兵,欧长波,等.益生菌发酵饲料对仔猪生长和免疫功能影响的研究进展[J].中国畜牧兽医,2017,44(2):476-481ZHANG X L,WEI X B,OU C B,et al. Research progress of probiotics fermented feed effects on growth performance and immune function in piglets[J]. China Animal Husbandry&Veterinary Medicine,2017,44(2):476-481
[12]吴妍妍,张文举,聂存喜,等.棉粕源发酵饲料产物中有机酸、B族维生素和游离氨基酸的研究[J].粮食与饲料工业,2013,12(5):46-50WU Y Y, ZHANG W J, NIE C X, et al. Study on organic acid, vitamin B and free amino acid in fermented feed products of cottonseed meal[J]. Cereal&Feed Industry, 2013,12(5):46-50
[13]陈来,冷向军,李小勤,等.优化高效液相色谱法测定棉籽粕中游离棉酚的含量[J].动物营养学报,2012,24(7):1320-1328CHEN L,LENG X J,LI X Q,et al. Optimized HPLC. method for determining free gossypol content in cottonseed meal[J]. Chinese Journal of Animal Nutrition. 2012,24(7):1320-1328
[14]GB 5009. 148—2014,食品安全国家标准植物性食品中游离棉酚的测定[S]GB 5009. 148—2014, National standard for food safety determination of free gossypol in vegetable foods[S]
[15]乔晓艳,蔡国林,陆健.微生物发酵改善棉粕饲用品质的研究[J].中国油脂,2013,38(5):30-34QIAO X Y, CAI G L, LU J. Improvement of feeding quality of cottonseed meal by microbial fermentation[J]. China Oils and Fats,2013,38(5):30-34
[16]杨霞,翁晓燕,郭建林,等.棉酚降解菌株的分离、筛选及鉴定[J].棉花学报,2010,22(6):539-546YANG X.WENG X Y.GUO J L,et al. Screening and identification of gossypol-degraded strains isolated from a soil microcosm[J]. Cotton Science,2010,22(6):539-546
[17]闫亚婷.固态发酵玉米条件的优化及营养物质变化的比较研究[D].雅安:四川农业大学,2010YAN Y T. Optimiation of solid-state fermentation of corn meal and analyze the change of nutritional ingredient[D].Yaan:Sichuan Agriculture University,2010
[18]刘颖,张彬彬,孙冰玉,等.枯草芽孢杆菌高产中性蛋白酶发酵条件的优化[J].食品科学,2014, 35(13):166-170LIU Y, ZHANG B B,SUN B Y,et al. Optimization of fermentation conditions for the production of neutral protease by Bacillus subtilis[J]. Food Science, 2014, 35(13):166-170
[19]胡晓清,褚召娟,张继祥,等.酵母生物脱毒的研究进展[J].现代食品科技,2017,33(12):272-277HU X Q,CHU S J, ZHANG J X.et al. Advances on Yeast biological detoxification[J]. Modern Food Science and Technology,2017,33(12):272-277
[20]程福亮,梁瑾,范群平,等.产朊假丝酵母菌发酵降解棉酚的效果[J].饲料广角,2012,14:22-25CHENG F L, LIANG J,FAN Q P,et al. The effect of C.andidautilis on the degradation of gossypol[J]. Feed China,2012,14:22-25
[21]周慧林,梅雪臣,缪礼鸿.一株降解棉酚的酵母菌培养特征及发酵效果分析[J].武汉轻工大学学报,2014,33(1):14-16ZHOU H L, MEI X C, LIAO L H. Analysis of the cultural characteristics and the effect of fermentation of yeast degrading gossypol[J]. Journal of Wuhan Polytechnic University,2014,33(1):14-16
[22]陈生琴,陈丽娟,程茂基,等.高效降解棉酚菌株的选育及发酵棉粕最佳工艺的研究[J].中国油脂,2016,41(4):19-24CHEN S Q, CHEN L J, CHENG M J, et al. Screening of high efficient gossypol-degrading strain and optimization of cottonseed meal[J]. China Oils and Fats.2016.41(4):19-24
[23]张文举,许梓荣,孙建义,等.假丝酵母ZD-3固体发酵对棉子饼脱毒的效果研究[J].棉花学报,2006,18(5):259-263ZHANG W J,XU Z R.SUNJ Y.et al. Effect of solid substrate fermentation of C. tropicalis ZD-3 on detoxif ication of cottonseed meal[J]. Cotton Science, 2006, 18(5):259-263
[24]王晓伟,高鹏飞,姚国强,等.乳酸菌发酵饲料的优势及其在畜禽养殖中的应用[J].粮食与饲料工业,2015,12(7):47-50WANG X W, GAO P F, YAO G Q, et al. Advantages of Lactic acid bacteria fermented feed and its application in livestock and poultry breeding[J]. Cereal&Feed Industry,2015,12(7):47-50
[25]刘建成,马燕,姚峻,等.饲用乳酸菌固态发酵棉籽粕产有机酸的研究[J].饲料与畜牧,2013(12):24-27LIU J C,MA Y,YAO J,et al. Study on solid-state fermen-tation of cottonseed meal to produce organic acids by Lactobacillus feed[J]. Feed and Animal Husbandry, 2013(12):24-27
[26]魏炳栋,党修利,邱玉朗,等.乳酸菌固态发酵酶解对豆粕、棉籽粕和菜籽粕粗蛋白质、pH、酸度及抗营养因子含量的影响[J].中国畜牧兽医,2014,41(11):107-114WEI B D,DANG X L,QIU Y L,et al. Effects of Lactic acid bacteria solid-state fermentation on crude protein, pH, acidity and antinutritional factor content of soybean meal, cottonseed meal and rapeseed meal[J]. China Animal Husbandry&Veterinary Medicine,2014,41(11):107-114
[27]诸葛斌,刘俊,方慧英,等.混菌发酵改良棉粕蛋白工艺及协同作用研究[J].中国生物工程杂志,2011,31(9):62-68ZHU G B,LIU J,FANG H Y,et al. Optimization of mixed fermentation on cottonseed meal and research on the synergistic effect of B. subtilis H1 and A. niger P1[J]. China BioTechnology,2011,31(9):62-68
[28]乔晓艳,蔡国林,陆健.微生物发酵改善棉粕饲用品质的研究[J].中国油脂,2013,38(5):30-34QIAO X Y, CAI G L, LU J. Improvement of feeding quality of cottonseed meal by microbial fermentation[J]. China Oils and Fats, 2013,38(5):30-34
[29]刘惠琴,田永强,徐力,等.混菌固态发酵棉粕工艺参数的优化[J].食品工业科技,2016,17(36):211-219LIU H Q, TIAN Y Q, XU L, et al. Optimization of the process parameters of cottonseed cake by solid-state fermentation with mixed culture[J]. Science and Technology of Food Industry,2016,17(36):211-219
[30]刘彩霞,郭学武,李玲玲,等.高产酯酿酒酵母与乳酸菌共发酵过程中的相互作用研究[J].现代食品科技,2017,33(7):79-84LIU C X.GUO X W,LI L L,et al. Interactions of high ester producing saccharomyces cerevisiae and Lactic acid bacteria during co-fermentation[J]. Modern Food Science and Technology,2017,33(7):79-84.
[2]亓秀晔,谢全喜,陈振,等.高效降解游离棉酚并改善棉籽粕营养品质的菌株筛选[J].动物营养学报,2017,29(9):3258-3266QI X Y,XIE Q X,CHEN Z,et al. Screening of high efficient free gossypol-degrading and improving cottonseed meal nutritive quality strains[J]. Chinese Journal of Animal Nutrition,2017,29(9):3258-3266
[3]陈生琴,陈丽娟,程茂基.高效降解棉酚菌株的选育及发酵棉粕最佳工艺的研究[J].中国油脂,2016.41(4):19-24CHEN S Q,CHEN L J,CHENG M J,et al. Screening of high efficient gossypol-degrading strain andoptimization of cottonseed meal[J]. China Oils and Fats,2016,41(4):19-24
[4]LORDELO M M, CALHOUN M C, DALE N M, et al. Relative toxicity of gossypol enantiomers in laying and broiler breeder hens[J]. Poultry Science,2007,86:582-590
[5]吕云峰,王修启,赵青余,等.棉酚在饲料中安全限量及畜产品中残留研究进展[J].中国农学通报,2010,26(24):1-5LYU Y F,WANG X Q,ZHAO Q Y,et al. Research situation on gossypol safety limit in feed and gossypol residues in livestock product[J]. Chinese Agricultural Science Bulletin,2010,26(24):1-5
[6]YANG X, SUN J Y, GUO J L, et al. Identification and proteomic analysis of a novel gossypol-degrading fungal strain[J]. Journal of the Science of Food Agriculture, 2012,92(4):943-951
[7]SUN H, TANG J W, YAO X H, et al. Improvement of the nutritional quality of cottonseed meal by Bacillus subtilis and the addition of papain[J]. International Journal of Agriculture and Biology, 2012, 14(4):563-568
[8]DAI C H,ZHANG L P,MA H L. Ultrasound-assisted detoxification of free gossypol from cottonseed meal[J]. Journal of Food Process Engineering, 2015. 7,https://doi.org/10. 1111/jfpe. 12265
[9]张煜,石常友,王成,等.菌酶协同发酵改善玉米-豆粕型饲料营养价值的研究[J].中国粮油学报,2018,33(3):70-77ZHANG Y,SHI C Y,WANG C,et al. Improving nutrition value of maize-soybean meal based diet by microbial fermenta-tion combined with multiple-enzymes[J]. Journal of the Chinese Cereals and Oils Association,2018,33(3):70-77
[10]张连慧,熊小辉,惠菊,等.发酵豆粕及其在动物养殖行业中的应用研究进展[J].中国油脂,2017,42(3):108-112ZHANG L H,XIONG X H,HUI J,et al. Progress in fermented soybean meal and application in animal breeding industry[J]. China Oils and Fats, 2017,42(3):108-112
[11]张秀林,魏小兵,欧长波,等.益生菌发酵饲料对仔猪生长和免疫功能影响的研究进展[J].中国畜牧兽医,2017,44(2):476-481ZHANG X L,WEI X B,OU C B,et al. Research progress of probiotics fermented feed effects on growth performance and immune function in piglets[J]. China Animal Husbandry&Veterinary Medicine,2017,44(2):476-481
[12]吴妍妍,张文举,聂存喜,等.棉粕源发酵饲料产物中有机酸、B族维生素和游离氨基酸的研究[J].粮食与饲料工业,2013,12(5):46-50WU Y Y, ZHANG W J, NIE C X, et al. Study on organic acid, vitamin B and free amino acid in fermented feed products of cottonseed meal[J]. Cereal&Feed Industry, 2013,12(5):46-50
[13]陈来,冷向军,李小勤,等.优化高效液相色谱法测定棉籽粕中游离棉酚的含量[J].动物营养学报,2012,24(7):1320-1328CHEN L,LENG X J,LI X Q,et al. Optimized HPLC. method for determining free gossypol content in cottonseed meal[J]. Chinese Journal of Animal Nutrition. 2012,24(7):1320-1328
[14]GB 5009. 148—2014,食品安全国家标准植物性食品中游离棉酚的测定[S]GB 5009. 148—2014, National standard for food safety determination of free gossypol in vegetable foods[S]
[15]乔晓艳,蔡国林,陆健.微生物发酵改善棉粕饲用品质的研究[J].中国油脂,2013,38(5):30-34QIAO X Y, CAI G L, LU J. Improvement of feeding quality of cottonseed meal by microbial fermentation[J]. China Oils and Fats,2013,38(5):30-34
[16]杨霞,翁晓燕,郭建林,等.棉酚降解菌株的分离、筛选及鉴定[J].棉花学报,2010,22(6):539-546YANG X.WENG X Y.GUO J L,et al. Screening and identification of gossypol-degraded strains isolated from a soil microcosm[J]. Cotton Science,2010,22(6):539-546
[17]闫亚婷.固态发酵玉米条件的优化及营养物质变化的比较研究[D].雅安:四川农业大学,2010YAN Y T. Optimiation of solid-state fermentation of corn meal and analyze the change of nutritional ingredient[D].Yaan:Sichuan Agriculture University,2010
[18]刘颖,张彬彬,孙冰玉,等.枯草芽孢杆菌高产中性蛋白酶发酵条件的优化[J].食品科学,2014, 35(13):166-170LIU Y, ZHANG B B,SUN B Y,et al. Optimization of fermentation conditions for the production of neutral protease by Bacillus subtilis[J]. Food Science, 2014, 35(13):166-170
[19]胡晓清,褚召娟,张继祥,等.酵母生物脱毒的研究进展[J].现代食品科技,2017,33(12):272-277HU X Q,CHU S J, ZHANG J X.et al. Advances on Yeast biological detoxification[J]. Modern Food Science and Technology,2017,33(12):272-277
[20]程福亮,梁瑾,范群平,等.产朊假丝酵母菌发酵降解棉酚的效果[J].饲料广角,2012,14:22-25CHENG F L, LIANG J,FAN Q P,et al. The effect of C.andidautilis on the degradation of gossypol[J]. Feed China,2012,14:22-25
[21]周慧林,梅雪臣,缪礼鸿.一株降解棉酚的酵母菌培养特征及发酵效果分析[J].武汉轻工大学学报,2014,33(1):14-16ZHOU H L, MEI X C, LIAO L H. Analysis of the cultural characteristics and the effect of fermentation of yeast degrading gossypol[J]. Journal of Wuhan Polytechnic University,2014,33(1):14-16
[22]陈生琴,陈丽娟,程茂基,等.高效降解棉酚菌株的选育及发酵棉粕最佳工艺的研究[J].中国油脂,2016,41(4):19-24CHEN S Q, CHEN L J, CHENG M J, et al. Screening of high efficient gossypol-degrading strain and optimization of cottonseed meal[J]. China Oils and Fats.2016.41(4):19-24
[23]张文举,许梓荣,孙建义,等.假丝酵母ZD-3固体发酵对棉子饼脱毒的效果研究[J].棉花学报,2006,18(5):259-263ZHANG W J,XU Z R.SUNJ Y.et al. Effect of solid substrate fermentation of C. tropicalis ZD-3 on detoxif ication of cottonseed meal[J]. Cotton Science, 2006, 18(5):259-263
[24]王晓伟,高鹏飞,姚国强,等.乳酸菌发酵饲料的优势及其在畜禽养殖中的应用[J].粮食与饲料工业,2015,12(7):47-50WANG X W, GAO P F, YAO G Q, et al. Advantages of Lactic acid bacteria fermented feed and its application in livestock and poultry breeding[J]. Cereal&Feed Industry,2015,12(7):47-50
[25]刘建成,马燕,姚峻,等.饲用乳酸菌固态发酵棉籽粕产有机酸的研究[J].饲料与畜牧,2013(12):24-27LIU J C,MA Y,YAO J,et al. Study on solid-state fermen-tation of cottonseed meal to produce organic acids by Lactobacillus feed[J]. Feed and Animal Husbandry, 2013(12):24-27
[26]魏炳栋,党修利,邱玉朗,等.乳酸菌固态发酵酶解对豆粕、棉籽粕和菜籽粕粗蛋白质、pH、酸度及抗营养因子含量的影响[J].中国畜牧兽医,2014,41(11):107-114WEI B D,DANG X L,QIU Y L,et al. Effects of Lactic acid bacteria solid-state fermentation on crude protein, pH, acidity and antinutritional factor content of soybean meal, cottonseed meal and rapeseed meal[J]. China Animal Husbandry&Veterinary Medicine,2014,41(11):107-114
[27]诸葛斌,刘俊,方慧英,等.混菌发酵改良棉粕蛋白工艺及协同作用研究[J].中国生物工程杂志,2011,31(9):62-68ZHU G B,LIU J,FANG H Y,et al. Optimization of mixed fermentation on cottonseed meal and research on the synergistic effect of B. subtilis H1 and A. niger P1[J]. China BioTechnology,2011,31(9):62-68
[28]乔晓艳,蔡国林,陆健.微生物发酵改善棉粕饲用品质的研究[J].中国油脂,2013,38(5):30-34QIAO X Y, CAI G L, LU J. Improvement of feeding quality of cottonseed meal by microbial fermentation[J]. China Oils and Fats, 2013,38(5):30-34
[29]刘惠琴,田永强,徐力,等.混菌固态发酵棉粕工艺参数的优化[J].食品工业科技,2016,17(36):211-219LIU H Q, TIAN Y Q, XU L, et al. Optimization of the process parameters of cottonseed cake by solid-state fermentation with mixed culture[J]. Science and Technology of Food Industry,2016,17(36):211-219
[30]刘彩霞,郭学武,李玲玲,等.高产酯酿酒酵母与乳酸菌共发酵过程中的相互作用研究[J].现代食品科技,2017,33(7):79-84LIU C X.GUO X W,LI L L,et al. Interactions of high ester producing saccharomyces cerevisiae and Lactic acid bacteria during co-fermentation[J]. Modern Food Science and Technology,2017,33(7):79-84.