大豆渣提取L-阿拉伯糖及辅助玉米秸秆酶解液发酵产乙醇的研究
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摘要
对大豆渣组成成分进行分析,并对其L-阿拉伯糖的提取条件进行单因素试验优化,并通过在发酵过程中补加酸解大豆渣(ASD)或酶解大豆渣(ESD),以酿酒酵母(Saccharomyces cerevisiae)LF1对稀酸预处理玉米秸秆酶解液(EHCS)进行分批及补料分批发酵产乙醇。结果表明,葡萄糖、半乳糖、L-阿拉伯糖和蛋白质分别约占大豆渣生物质干质量的16.4%、18.8%、10.5%和24.1%;L-阿拉伯糖最佳提取条件为大豆渣与0.5mol/L稀硫酸的料液比1∶5(g∶mL)、提取温度100℃、提取时间45min,在此最佳提取条件下,L-阿拉伯糖含量为21.0g/L;EHCS发酵补料ASD或ESD,乙醇产量均可达到110g/L以上,比分批发酵乙醇产量提高了37%,且ESD和尿素组合使用可以替代发酵氮源。
The composition of soybean dreg was analyzed and the extraction conditions of L-arabinose were optimized by single factor tests. Through feeding acidolysis soybean dregs(ASD) or enzymolysis soybean dregs(ESD) during fermentation,ethanol was produced by batch and fed-batch fermentation of Saccharomyces cerevisiae LF1 with the enzymatic hydrolysate of corn stover(EHCS) pretreated with dilute acid. The results showed that the glucose, galactose, L-arabinose and protein accounted for 16.4%, 18.8%, 10.5% and 24.1% of the dry mass of soybean dreg, respectively. The optimal extraction conditions of L-arabinose were the soybean dregs to 0.5 mol/L dilute sulfuric acid ratio 1∶5(g∶ml), extraction temperature 100 ℃ and time 45 min, and the L-arabinosecontent was 21.0 g/L under the optimal condition. When the EHCS was fermented by feeding ASD or ESD, the ethanol production could reach above 110 g/L, which increased by 37% compared with the batch fermentation, and the combination of ESD and urea could replace fermentation nitrogen sources.
The composition of soybean dreg was analyzed and the extraction conditions of L-arabinose were optimized by single factor tests. Through feeding acidolysis soybean dregs(ASD) or enzymolysis soybean dregs(ESD) during fermentation,ethanol was produced by batch and fed-batch fermentation of Saccharomyces cerevisiae LF1 with the enzymatic hydrolysate of corn stover(EHCS) pretreated with dilute acid. The results showed that the glucose, galactose, L-arabinose and protein accounted for 16.4%, 18.8%, 10.5% and 24.1% of the dry mass of soybean dreg, respectively. The optimal extraction conditions of L-arabinose were the soybean dregs to 0.5 mol/L dilute sulfuric acid ratio 1∶5(g∶ml), extraction temperature 100 ℃ and time 45 min, and the L-arabinosecontent was 21.0 g/L under the optimal condition. When the EHCS was fermented by feeding ASD or ESD, the ethanol production could reach above 110 g/L, which increased by 37% compared with the batch fermentation, and the combination of ESD and urea could replace fermentation nitrogen sources.
引文
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[2]赵影,韩建春,郑环宇,等.豆渣深加工及综合利用的研究现状[J].大豆科学,2013,32(4):555-560.
[3]董英.大豆渣营养价值及其综合利用[J].粮食与油脂,2001(12):41-42.
[4]桂雨豪.水压热法制备水溶性大豆多糖与其应用研究[D].广州:华南理工大学,2016.
[5]KAATS G R,KEITH S C,KEITH P L,et al.A combination of L-arabinose and chromium lowers circulating glucose and insulin levels after an acute oral sucrose challenge[J].Nutr J,2011,10(1):42.
[6]郑敏.玉米皮中L-阿拉伯糖的制备工艺的研究[D].天津:天津科技大学,2012.
[7]袁其朋,王乐,范晓光,等.一种清洁高效的制备木糖与L-阿拉伯糖的生产工艺:CN102286571B[P].2014-04-02.
[8]GAO J,FENG H,YUAN W,et al.Enhanced fermentative performance under stresses of multiple lignocellulose-derived inhibitors by overexpression of a typical 2-Cys peroxiredoxin from Kluyveromyces marxianus[J].Biotech Biofuels,2017,10(1):79.
[9]LI H,SHEN Y,WU M,et al.Engineering a wild-type diploid Saccharomyces cerevisiae strain for second-generation bioethanol production[J].Bioresources Bioproc,2016,3(1):51.
[10]ZHANG H,LIU G,ZHANG J,et al.Fermentative production of high titer gluconic and xylonic acids from corn stover feedstock by Gluconobacter oxydans and techno-economic analysis[J].Bioresource Technol,2016,219:123-131.
[11]WANG G,THOMPSON M R,LIU Q.Controlling the moisture absorption capacity in a fiber-reinforced thermoplastic starch using sodium trimetaphosphate[J].Ind Crop Prod,2012,36(1):299-303.
[12]中华人民共和国国家卫生和计划生育委员会.GB 5009.4-2010食品中灰分的测定[S].北京:中国标准出版社,2010.
[13]苗霖兴,苗佳.大豆粗脂肪含量测定中影响因素的分析[J].粮食科技与经济,2011,36(3):42-42.
[14]韦芳三,李纯厚,戴明,等.索氏提取法测定海洋微藻粗脂肪含量及其优化方法的研究[J].上海海洋大学学报,2011,20(4):619-623.
[15]中华人民共和国国家卫生和计划生育委员会,国家食品药品监督管理总局.GB 5009.6-2016食品中脂肪的测定[S].北京:中国标准出版社,2008.
[16]HYMAN D,SLUITER A,CROCKER D,et al.Determination of acid soluble lignin concentration curve by UV-vis spectroscopy[J].Natl Renew Energ Lab Anal Proced,2008:1-13.
[17]谭丽萍.油棕空果壳预处理、糖化发酵生产乙醇及预处理机制的研究[D].济南:山东大学,2015.
[18]中华人民共和国国家卫生和计划生育委员会,国家食品药品监督管理总局.GB 5009.5-2016食品中蛋白质含量的测定[S].北京:中国标准出版社,2010.
[19]CHROM Y′V,VINKL魣RKOV魣B,譒PRONGL L,et al.The Kjeldahl method as a prim ary reference procedure for total protein in certified reference materials used in clinical chemistry.I.A review of Kjeldahl methods adopted by laboratory medicine[J].Crit Rev Anal Chem,2015,45(2):106-111.
[20]齐建双,铁双贵,韩小花,等.氨基酸分析仪法快速测定玉米籽粒中赖氨酸含量[J].中国农学通报,2014,30(30):199-202.
[21]黄晓荣,张良晓,李培武,等.黑芝麻和白芝麻中氨基酸组成的比较研究[J].中国油料作物学报,2017,39(1):123.
[22]中华人民共和国国家卫生和计划生育委员会,国家食品药品监督管理总局.GB 5009.9-2016食品中淀粉的测定[S].北京:中国标准出版社,2016.
[23]WANG C,BAO X,LI Y,et al.Cloning and characterization of heterologous transporters in Saccharomyces cerevisiae and identification of important amino acids for xylose utilization[J].Metab Eng,2015,30:79-88.
[24]SAHOO D,UMMALYMA S B,OKRAM A K,et al.Effect of dilute acid pretreatment of wild rice grass(Zizania latifolia)from Loktak Lake for enzymatic hydrolysis[J].Bioresource technol,2018,253:252-255.
[25]LI H,SCHMITZ O,ALPER H S.Enabling glucose/xylose co-transport in yeast through the directed evolution of a sugar transporter[J].Appl Microbiol Biot,2016,100(23):10215-10223.
[26]WEI S,LIU Y,WU M,et al.Disruption of the transcription factors Thi2p and Nrm1p alleviates the post-glucose effect on xylose utilization in Saccharomyces cerevisiae[J].Biotech Biofuels,2018,11(1):112.