热碱法提取异养小球藻蛋白质工艺优化
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摘要
在异养小球藻提油之前提取蛋白质,为降低产油成本奠定基础。以蛋白质提取率为指标,使用热碱法提取小球藻中的蛋白质,首先采用单因素试验确定提取过程中主要影响因素的最佳水平,然后利用正交试验对主要因素的相互作用进行研究。结果表明,小球藻在Na OH添加量为20%(质量分数),液料比为50∶1,提取时间为60 min,提取温度为55℃的条件下蛋白质的提取率最高,为55. 24%。热碱法提取异养小球藻蛋白,提取率高、操作简单、成本低,具有较好的工业化应用前景,为异养小球藻产油技术的经济最大化提供了参考。
Protein extraction from Heterotrophic chlorella before extracting oil can reduce its oil extraction cost. In this study,the protein extraction rate was used as an index,the hot alkali method was used to extract protein from Heterotrophic chlorella. Single factor test was used to determine the best level of main factors in the extraction process.Then the interaction of main factors was studied by orthogonal test. The results showed that the best extracting conditions were: NaOH 20%,solid-liquid ratio 50 ∶ 1,extraction time 60 min and extraction temperature 55 ℃. Under these conditions,the protein extraction rate of Heterotrophic chlorella was the highest of 55. 24%. By hot alkali method,the extraction rate was high,the operation was simple and the cost was low. It has a good potential in industrial application. This study provides a reference for an economic maximization of Heterotrophic chlorella oil production.
Protein extraction from Heterotrophic chlorella before extracting oil can reduce its oil extraction cost. In this study,the protein extraction rate was used as an index,the hot alkali method was used to extract protein from Heterotrophic chlorella. Single factor test was used to determine the best level of main factors in the extraction process.Then the interaction of main factors was studied by orthogonal test. The results showed that the best extracting conditions were: NaOH 20%,solid-liquid ratio 50 ∶ 1,extraction time 60 min and extraction temperature 55 ℃. Under these conditions,the protein extraction rate of Heterotrophic chlorella was the highest of 55. 24%. By hot alkali method,the extraction rate was high,the operation was simple and the cost was low. It has a good potential in industrial application. This study provides a reference for an economic maximization of Heterotrophic chlorella oil production.
引文
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[43]陈艺煊,刘晓艳,吴林秀,等.蛋白核小球藻多糖的酶解辅助提取及抗氧化活性[J].福建农业学报,2016,31(5):508-514.
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[3]刘彩霞.小球藻(Chlorella vulgaris)在不同浓度Fe~(3+)培养下的蛋白质组学初步研究[D].北京:中国科学院大学,2016.
[4]庄秀园,黄英明,张道敬,等.小球藻高附加值生物活性物质“小球藻热水提取物”的研究现状与展望[J].生物工程学报,2015,31(1):24-42.
[5]薄香兰,窦勇,张文慧,等.小球藻生长因子的研究进展[C]//第一届微藻与水族高峰论坛论文摘要集.天津:天津现代晨辉科技集团有限公司,2017.
[6]何扩,张秀媛,李玉锋.小球藻破壁技术及其藻片研制[J].食品工业科技,2006(2):32-33.
[7] LIN P Y,TSAI C T,CHUANG W L,et al. Chlorella sorokiniana induces mitochondrial-mediated apoptosis in human non-small cell lung cancer cells and inhibits xenograft tumor growth in vivo[J]. Bmc Complementary&Alternative Medicine,2017,17(1):88.
[8]韩士群,张振华,刘海琴.小球藻生长因子对免疫功能的影响[J].中国生化药物杂志,2004,25(1):5-7.
[9] ZHANG Z D,LIANG K,LI K,et al. Chlorella vulgaris induces apoptosis of human non-small cell lung carcinoma(NSCLC)cells.[J]. Medicinal Chemistry,2017,13(999):1.
[10] KIM Y H,HWANG Y K,KIM Y Y,et al. Effect of chlorella growth factor on the proliferation of human skin keratinocyte[J]. Biomedical Science Letters,2002,8:229-234.
[11]梅帅,赵凤敏,曹有福,等.三种小球藻的蛋白质营养价值评价[J].营养学报,2014,36(5):505-507.
[12]葛珍珍.小球藻的筛选、培养和产油脂研究[D].无锡:江南大学,2012.
[13]李明,刘红.富油微藻培养的研究进展[C]//2010中国可再生能源科技发展大会论文集.北京:中国能源协会,2015:1 028-1 032.
[14]孔维宝,汪洋,杨红,等.不同营养方式对普通小球藻生长代谢及生化组分的影响[J].微生物学报,2015,55(3):299-310.
[15]刘世名.小球藻(Chlorella Vulgaris)高密度异养培养[D].广州:华南理工大学,1999.
[16]马千然,汪苹.发酵罐高密度异养培养小球藻的研究[J].广东化工,2013,40(4):15-17.
[17]吴正云.小球藻异养培养的动力学分析与优化[D].上海:上海交通大学,2007.
[18]毕生雷,刘钺,杜风光,等.异养小球藻制生物柴油技术研究进展[C]//全国微藻生物能源技术创新交流研讨会暨新产品、新工艺、新设备展示与合作对接会.杭州:中国高科技产业化研究会,2016:13-19.
[19]吴庆余,熊伟.一种从自养到异养两步培养小球藻生产生物柴油的方法:CN,CN101280328[P]. 2008.
[20]李双双.小球藻的培养及产油条件研究[D].杭州:浙江农林大学,2014.
[21]黄俊远.从小球藻中提取油脂、蛋白质和多糖的研究[D].北京:北京化工大学,2014.
[22]林芃,刘艳,杨海波.对小球藻蛋白质的提取及分离的研究[J].大连大学学报,2002,23(4):70-73.
[23]王晓琴.小球藻抗肿瘤多肽的分离及其微囊化研究[D].广州:华南理工大学,2012.
[24]汤永强,范宇,范伟权.小球藻压力破壁及核苷酸、蛋白质、多糖、藻干粉的制取方法:CN101053577[P]. 2007.
[25] LI Y Q,HAN F X,XU H,et al. Potential lipid accumulation and growth characteristic of the green alga Chlorella with combination cultivation mode of nitrogen(N)and phosphorus(P)[J]. Bioresour Technol,2014,174:24-32.
[26]纪雁.利用味精废水培养普通小球藻以及养藻废水的生物强化处理[D].济南:山东大学,2014.
[27]魏东,张会贞,陈娇敏.优化营养方式强化蛋白核小球藻生物量及蛋白质和叶绿素生产[J].现代食品科技,2017(4):160-167.
[28] ZHU S,HUANG W,XU J,et al. Metabolic changes of starch and lipid triggered by nitrogen starvation in the microalga Chlorella zofingiensis[J]. Bioresource Technology,2014,152(1):292-298.
[29] ZHU S,WANG Y,SHANG C,et al. Characterization of lipid and fatty acids composition of Chlorella zofingiensis in response to nitrogen starvation[J]. Journal of Bioscience&Bioengineering,2015,120(2):205.
[30]刘四光.自养小球藻(Chlorella vulgaris)多糖提取、纯化、化学结构及抗菌活性研究[D].厦门:厦门大学,2017.
[31]李兴武,李元广,万俊芬,等.普通小球藻异养-光自养串联培养的研究[C]//中国生物工程学会第四次会员代表大会暨学术讨论会论文摘要集.北京:中国生物工程学会,2005.
[32]吴庆余,盛国英,傅家谟.自养与异养小球藻脂溶性化合物对比研究[J].植物生态学报(英文版),1993(11):849-858.
[33] CHEN C Y,YEH K L,CHANG H Y,et al. Strategies to improve oil/lipid production of microalgae in outdoor cultivation using vertical tubular-type photobioreactors☆[J]. Energy Procedia,2014,61(1):2 755-2 758.
[34]李双双.小球藻的培养及产油条件研究[D].杭州:浙江农林大学,2014.
[35]占晶晶.外源物质和培养策略对小球藻产油和净水能力的影响[D].北京:北京林业大学,2016.
[36] RUANGSOMBOON S. Effect of light,nutrient,cultivation time and salinity on lipid production of newly isolated strain of the green microalga,Botryococcus braunii KMITL2[J]. Bioresource Technology,2012,109(109):261.
[37]曹维. DHA藻油在食用油中的应用研究[D].武汉:武汉轻工大学,2015.
[38] RICHMOND A,HU Q,RICHMOND A,et al. Handbook of microalgal culture:Applied phycology and biotechnology[J]. Handbook of Microalgal Culture Applied Phycology&Biotechnology,2013:146-167.
[39] ABEDINI N H,MALEKZADEH M,JALILIAN F,et al.Effect of various carbon sources on biomass and lipid production of Chlorella vulgaris during nutrient sufficient and nitrogen starvation conditions.[J]. Bioresource Technology,2015,180:311-317.
[40]钟韵山,徐仰仓,荆柏林,等.小球藻破壁技术研究进展[J].食品研究与开发,2014(14):120-124.
[41]曲文娟,马海乐,张厚森.钝顶螺旋藻藻蓝蛋白的脉冲超声辅助提取技术[J].食品科技,2007,32(5):135-139.
[42]王晓琴,张学武.响应面法优化小球藻蛋白质提取工艺[J].现代食品科技,2012,28(3):300-303.
[43]陈艺煊,刘晓艳,吴林秀,等.蛋白核小球藻多糖的酶解辅助提取及抗氧化活性[J].福建农业学报,2016,31(5):508-514.
[44]刘光明,李林,.碱提酸沉法制备燕麦浓缩蛋白的工艺研究[J].湖南文理学院学报(自科版),2011,23(1):60-62.
[45]江怀真,刘天中,高莉丽,等.热蒸汽-Na OH法提取小球藻蛋白质的研究[J].安徽农业科学,2009,37(33):16 204-16 206.
[46]王喜刚,况楠,裴云生,等.脱脂豆粕中大豆分离蛋白提取工艺的研究[J].粮食与食品工业,2012,19(6):58-61.
[47]姜珊珊,王红艳,王乾,等.黑米蛋白提取工艺的优化[J].天然产物研究与开发,2016(1):142-146.
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