小球藻细胞匀浆制备方法探究

doi:10.19640/j.cnki.jtau.2021.03.014

天津农学院学报 ›› 2021, Vol. 28 ›› Issue (3): 65-68.doi: 10.19640/j.cnki.jtau.2021.03.014

小球藻细胞匀浆制备方法探究

王政润, 于建霖, 孔庆霞^通信作者

天津农学院水产学院,天津 300392

收稿日期:2020-01-15 出版日期:2021-09-30 发布日期:2021-10-07
通讯作者: 孔庆霞（1978—）,女,副教授,博士,主要从事环境毒理学方面的研究。E-mail：xiaokong3155@163.com。
作者简介:王政润（1995—）,男,硕士在读,主要从事环境毒理学方面的研究。E-mail：305179091@qq.com。
基金资助:
天津市水产产业技术体系尾水处理岗位（ITTFR2018047）; 天津市动植物抗性重点实验室开放基金项目（无编号）

Study on the preparation methods of homogenate chlorella cells

Wang Zhengrun, Yu Jianlin, Kong Qingxia^{Corresponding Author}

College of Fisheries, Tianjin Agricultural University, Tianjin 300392, China

Received:2020-01-15 Online:2021-09-30 Published:2021-10-07

摘要/Abstract

摘要： 以普通小球藻为受试生物,研究了液氮匀浆法、研磨匀浆法、机械匀浆法、反复冻融法等4种匀浆方法的破壁效率,同时使用4种方法制备的匀浆液测定SOD活力和GSH含量,旨在筛选出破碎效率高、方便快捷且测定生理生化指标准确的方法。试验结果表明,4种方法的破碎率分别为81.43%、100.00%、92.41%、56.54%,SOD活力分别为47.06、48.58、0、25.05 U/mgprot,其中研磨匀浆法与液氮破碎法的测定结果无显著差异（P>0.05）;GSH含量分别为34.42、24.52、87.37、38.75 mgGSH/gprot,研磨匀浆法、液氮破碎法、反复冻融法的测定结果无显著差异（P>0.05）,与机械匀浆法差异显著（P<0.05）。结果说明,研磨匀浆法和液氮破碎法的破碎率高,生化指标测定准确。但由于研磨匀浆法耗费人工成本过高,且无法处理大量样本,相较而言,在上述方法中采用液氮破碎法是制备小球藻细胞匀浆液最好的方法。

关键词: 普通小球藻, 匀浆方法, 细胞破碎率, 生化指标

Abstract: With Chlorella. vulgaris as the test organism, four homogenizate methods ,liquid nitrogen method, grinding method, mechanical method, and repeated freeze-thaw method were studied. The homogenation with the above methods was used to determined the SOD activity and GSH content, which aimed to select a method with high crushing efficiency, convenience, and accurate determination of physiological and biochemical indicators. The results showed that the crushing rates of the four methods were 81.43%, 100%, 92.41%, 56.54%, and the SOD activity were 47.06, 48.58, 0, 25.05 U/mgprot. There was no significant difference between liquid nitrogen method and grinding method（p>0.05）; GSH content were 34.42, 24.52, 87.37, 38.75 mgGSH/gprot; There was no significant difference apart of mechanical method（p>0.05）. The above results indicated that the grinding method and liquid nitrogen method have a high crushing rate, and accurate measurement of the biochemical indicators. However, grinding method consumed too much labor and cannot process a large number of samples, so the liquid nitrogen method was considered to be the best solution for preparing microalgal cell homogenation.

Key words: Chlorella. vulgaris, homogenate methods, crushing rate, biochemical indicators

中图分类号:

王政润, 于建霖, 孔庆霞. 小球藻细胞匀浆制备方法探究[J]. 天津农学院学报, 2021, 28(3): 65-68.

Wang Zhengrun, Yu Jianlin, Kong Qingxia. Study on the preparation methods of homogenate chlorella cells[J]. Journal of Tianjin Agricultural University, 2021, 28(3): 65-68.

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http://xuebao.tjau.edu.cn/CN/Y2021/V28/I3/65

参考文献

[1] 洪华嫦,周海云,蓝崇钰. 五氯酚对斜生栅藻的毒性效应研究[J]. 环境科学研究,2003(6):23-25.
[2] 陈香洁. TiO₂纳米颗粒在小球藻–大型蚤食物链中的传递与毒性效应[D]. 杭州:浙江大学,2019.
[3] LI M,LUO X,ZHANG X,et al.A common variant of the cardiomyopathy associated 5 gene(CMYA5)is associated with schizophrenia in Chinese population[J]. Schizophrenia Research,2011,129(2-3):217-219.
[4] 隋琪. 海洋酸化条件下纳米TiO₂颗粒对蛋白核小球藻的毒性效应[D]. 上海:上海海洋大学,2016.
[5] LI X,GAO Y,ZHOU X,et al.Nano–ZnO/regenerated cellulose films prepared by low–temperature hydrothermal decomposition of Zn₅(OH)₈Cl₂·H₂O from cellulose–ZnCl₂ aqueous solution[J]. Materials Letters,2019,245:82-85.
[6] COELHO D,LOPES P A,CARDOSO V,et al.Novel combination of feed enzymes to improve the degradation of Chlorella vulgaris recalcitrant cell wall[J]. Scientific Reports,2019,9(1):1-11.
[7] ZHANG C,TANG X H,YANG X Y.Overcoming the cell wall recalcitrance of heterotrophic Chlorella to promote the efficiency of lipid extraction[J]. Journal of Cleaner Production,2018,198:1224-1231.
[8] 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,2014,35(14):120-124.
[9] ZHANG W D,WANG J F,WANG J L,et al.Attached cultivation of Haematococcus pluvialis for astaxanthin production[J]. Bioresource Technology,2014,158:329-335.
[10] PARK J Y,LEE K,CHOI S A,et al.Sonication–assisted homogenization system for improved lipid extraction from Chlorella vulgaris[J]. Renewable Energy,2015,79(SI):3-8.
[11] DERAKHSHAN M V,NASERNEJAD B,DADVAR M,et al.Pretreatment and kinetics of oil extraction from algae for biodiesel production[J]. Asia–Pacific Journal of Chemical Engineering,2014,9(5):629-637.
[12] CHENG J,SUN J,HUANG Y,et al.Dynamic microstructures and fractal characterization of cell wall disruption for microwave irradiation–assisted lipid extraction from wet microalgae[J]. Bioresource Technology,2013,150:67-72.
[13] CONCAS A,PISU M,CAO G.Disruption of microalgal cells for lipid extraction through Fenton reaction:Modeling of experiments and remarks on its effect on lipids composition[J]. Chemical Engineering Journal,2015,263:392-401.
[14] PARK J Y,OH Y K,LEE J S,et al.Acid–catalyzed hot–water extraction of lipids from Chlorella vulgaris[J]. Bioresource Technology,2014,153:408-412.
[15] MENDES–PINTO M M,RAPOSO M,BOWEN J,et al. Evaluation of different cell disruption processes on encysted cells of Haematococcus pluvialis:effects on astaxanthin recovery and implications for bio–availability[J]. Journal of Applied Phycology,2001,13(1):19-24.
[16] CHEN C Y,BAI M D,CHANG J S.Improving microalgal oil collecting efficiency by pretreating the microalgal cell wall with destructive bacteria[J]. Biochemical Engineering Journal,2013,81:170-176.
[17] 韩继刚,刘平芳,康明,等. 小球藻病毒PBCV–1特异性溶壁酶(Lysin)的溶壁活性[J]. 微生物学报,2002(3):354-358.
[18] 王崔平. 活性氧自由基的检测及其清除研究[D]. 北京:首都师范大学,2007.
[19] 刘波,王庆奎,邢克智,等. 当归多糖除蛋白质前后抗氧化活性比较[J]. 天津农学院学报,2017,24(3):46-49.
[20] 齐金秋. 纳米氧化铜对莱茵衣藻的胁迫效应研究[D]. 泰安:山东农业大学,2016.
[21] 孙虹豆,王贝贝,张芝蕊,等. 两种地被植物对干旱胁迫的生理生化响应[J]. 天津农学院学报,2017,24(4):26-29.
[22] 刘源. 三种纳米氧化物对两种不同方式卤虫无节幼体的生物毒性效应研究[D]. 南京:南京农业大学,2016.
[23] 李琦,张兰威,韩雪,等. 破壁方法对嗜热链球菌SP1.1胞内乳糖代谢关键酶活性的影响及其条件优化[J]. 食品科学,2011,32(9):183-187.
[24] 袁勤生. 超氧化物歧化酶[M]. 上海:华东理工大学出版社,2009.

小球藻细胞匀浆制备方法探究

Study on the preparation methods of homogenate chlorella cells

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