产油单针藻的最适培养基筛选及产油特性提升

doi:10.11924/j.issn.1000-6850.casb2022-0048

中国农学通报 ›› 2022, Vol. 38 ›› Issue (27): 157-164.doi: 10.11924/j.issn.1000-6850.casb2022-0048

所属专题：生物技术

• 生物质能源 • 上一篇

产油单针藻的最适培养基筛选及产油特性提升

马思琦¹^,²(), 李畅¹^,², 樊阳¹^,², 孙莹¹^,², 尹紫良¹^,², 林宜萌¹^,²(), 葛菁萍¹^,²

1.黑龙江大学农业微生物技术教育部工程研究中心,哈尔滨 150500
2.黑龙江大学生命科学学院/黑龙江省普通高校微生物重点实验室,哈尔滨 150080

收稿日期:2022-01-26 修回日期:2022-05-02 出版日期:2022-10-05 发布日期:2022-09-21
通讯作者: 林宜萌
作者简介:马思琦,女,2000年出生,黑龙江鸡西人,本科,研究方向：微生物资源挖掘与利用。通信地址：150080 黑龙江省哈尔滨市南岗区学府路74号,Tel：0451-86609134,E-mail： 1015087387@qq.com。
基金资助:
黑龙江省自然科学基金项目“促进微藻产油的藻菌共培养系统建立及特征研究”(LH2020C089);国家自然科学基金项目“乙酸溢流与群体感应的协同效应对副干酪乳杆菌HD1.7种群稳定性影响机制的探究”(32071519);黑龙江省高校基本科研业务费黑龙江大学专项资金“单针藻HDMA-01在高铵条件下的响应特征与除铵潜能”(2021-KYYWF-0014)

Lipid-producing Monoraphidium sp.: The Best Medium Screening and Lipid-producing Property Enhancement

MA Siqi¹^,²(), LI Chang¹^,², FAN Yang¹^,², SUN Ying¹^,², YIN Ziliang¹^,², LIN Yimeng¹^,²(), GE Jingping¹^,²

1. Engineering Research Center of Agricultural Microbiology Technology, Ministry of Education, Heilongjiang University, Harbin 150500
2. Key Laboratory of Microbiology, College of Heilongjiang Province, School of Life Sciences, Heilongjiang University, Harbin 150080

Received:2022-01-26 Revised:2022-05-02 Online:2022-10-05 Published:2022-09-21
Contact: LIN Yimeng

摘要/Abstract

摘要：

为了研究培养基种类及成分对单针藻产油特性的影响,充分挖掘单针藻产油潜能,以Monoraphidium sp. HDMA-11为供试藻株,以细胞数监测、干重法和尼罗红染色法检测了BG-11培养基、BBM培养基、TAP培养基、D1培养基和SE培养基对藻株生长速度、生物量及含油量的影响,并对藻株可利用的碳源、氮源种类及浓度进行筛选与优化。结果表明,TAP培养基可极大缩短藻株培养周期,使藻株生物量生产率提升了127%,油脂生产率提升了117%。当乙酸添加量为2 mg/L,氯化铵浓度为750 mg/L,藻株的产油指标进一步提高。优化后藻株的生物量生产率为70 mg/(L·d),较初始培养条件提升了218%,优化后藻株的油脂生产率为315 RFU/(L·d),较初始培养条件提升了116%。本研究优化了Monoraphidium sp. HDMA-11的培养条件,明确了乙酸和氯化铵浓度对藻株生长特性及产油指标的影响,为进一步挖掘藻株产油潜能提供了数据支持。

关键词: 单针藻, 培养基, 产油特性, 生物量, 油脂含量

Abstract:

To investigate the effects of medium type and composition on the lipid-producing properties of Monoraphidium sp. HDMA-11 and fully improve the lipid production potential of the strain, the growth, biomass and lipid content of HDMA-11 were examined using cell counting, dry cell weight and Nile red staining methods in BG-11, BBM, TAP, D1 and SE media. The type and concentration of carbon and nitrogen sources were then optimized to improve the lipid production characteristics of the strain. Results showed that the TAP medium could greatly shorten the culture period of the strain and increase its biomass productivity by 127% and lipid productivity by 117%. The lipid-producing indexes of the strain were further improved when 2 mL/L of acetic acid and 750 mg/L of ammonium chloride were added. The productivity of biomass and lipid of the strain after being improved were 70 mg/(L·d) and 315 RFU/(L·d) respectively, which was 218% and 116% higher than those of the initial levels. This work optimized the culture conditions of HDMA-11, elucidated the effects of acetic acid and ammonium chloride on the growth and lipid-producing indexes, and provided data basis for further exploration of the lipid-producing potential of the strain.

Key words: Monoraphidium sp., medium, lipid-producing properties, biomass, lipid content

中图分类号:

S917

马思琦, 李畅, 樊阳, 孙莹, 尹紫良, 林宜萌, 葛菁萍. 产油单针藻的最适培养基筛选及产油特性提升[J]. 中国农学通报, 2022, 38(27): 157-164.

MA Siqi, LI Chang, FAN Yang, SUN Ying, YIN Ziliang, LIN Yimeng, GE Jingping. Lipid-producing Monoraphidium sp.: The Best Medium Screening and Lipid-producing Property Enhancement[J]. Chinese Agricultural Science Bulletin, 2022, 38(27): 157-164.

图/表 8

图1. Monoraphidium sp. HDMA-11在不同培养基中的生长曲线

图2. Monoraphidium sp. HDMA-11在不同培养基中的生物量、油脂含量 1. BG-11培养基;2. BBM培养基;3. TAP培养基;4. D1培养基;5. SE培养基

图3. 碳源种类对Monoraphidium sp. HDMA-11生物量(A)和油脂含量(B)的影响 1.果糖;2. 甘露醇;3. 麦芽糖;4. 乙酸;5. 甘油;6.乙酸钠;7.葡萄糖;8.蔗糖

图4. 氮源种类对Monoraphidium sp. HDMA-11生物量(A)和油脂含量(B)的影响 1. 硝酸铵;2. 硝酸钠;3. 氯化铵;4. 蛋白胨;5. 酵母提取物;6. 尿素

图5. 氮源种类对Monoraphidium sp. HDMA-11生长中OD680的影响

图6. 乙酸浓度对Monoraphidium sp. HDMA-11生物量(A)和油脂含量(B)的影响 1. 0 mL/L;2. 1 mL/L;3. 2 mL/L;4. 4 mL/L;5. 8 mL/L

图7. 氯化铵浓度对Monoraphidium sp. HDMA-11生物量(A)和油脂含量(B)的影响 1. 125 mg/L;2. 375 mg/L;3. 750 mg/L;4. 1500 mg/L;5. 3000 mg/L

培养基		生物量/(mg/L)	生物量生产率/[mg/(L·d)]	油脂含量/RFU	油脂生产率/[RFU/(L·d)]
BG-11		490	22	5.3	118
TAP	优化前	301	50	5.1	256
TAP	优化后	419	70	4.5	314

表1. HDMA-11在优化培养条件前后的生物量、生物量生产率、油脂含量和油脂生产率

参考文献 26

[1]	SHUBA E S, KIFLE D. Microalgae to biofuels: ‘Promising’ alternative and renewable energy, review[J]. Renewable & sustainable energy reviews, 2018, 81:743-755.
[2]	WANG H, PENG X, ZHANG H, et al. Microorganisms-promoted biodiesel production from biomass: a review[J]. Energy conversion and management, 2021, 12: 100137.
[3]	KUMAR V, JAISWAL K K, VERMA M, et al. Algae-based sustainable approach for simultaneous removal of micropollutants, and bacteria from urban wastewater and its real-time reuse for aquaculture[J]. Science of the total environment, 2021, 774(20):145556.
[4]	SITEPU E K, HEIMANN K, RASTON C L, et al. Critical evaluation of process parameters for direct biodiesel production from diverse feedstock[J]. Renewable and sustainable energy reviews, 2020, 123:109762.
[5]	BOROWITZKA M A. High-value products from microalgae-their development and commercialisation[J]. Journal of applied phycology, 2013, 25(3):743-756. doi: 10.1007/s10811-013-9983-9 URL
[6]	ROBERTSON R, GUIHENEUF F, SCHMID M, et al. Algae-derived polyunsaturated fatty acids: implications for human health[M]. 2013,7x10-(NBC-C).
[7]	BENEDETTI S, BENVENUTI F, PAGLIARANI S, et al. Antioxidant properties of a novel phycocyanin extract from the blue-green alga aphanizomenon flos-aquae[J]. Life sciences, 2004, 75(19): 2353-2362. doi: 10.1016/j.lfs.2004.06.004 URL
[8]	STOLZ P. Manufacturing microalgae for skin care[J]. Cosmetics toiletries, 2005,120.
[9]	LIU J, MAO X, ZHOU W, et al. Simultaneous production of triacylglycerol and high-value carotenoids by the astaxanthin-producing oleaginous green microalga Chlorella zofingiensis[J]. Bioresoure technology, 2016, 214:319-327. doi: 10.1016/j.biortech.2016.04.112 URL
[10]	GANESAN R, MANIGANDAN S, SAMUEL M S, et al. A review on prospective production of biofuel from microalgae[J]. Biotechnology reports, 2020, 27: e00509.
[11]	HAN F, PEI H, HU W, et al. Optimization and lipid production enhancement of microalgae culture by efficiently changing the conditions along with the growth-state[J]. Energy conversion & management, 2015, 90(1):315-322.
[12]	ZHAO Y, YUE C, GENG S, et al. Role of media composition in biomass and astaxanthin production of Haematococcus pluvialis under two-stage cultivation[J]. Bioprocess and biosystems engineering, 2019, 42(4):593-602. doi: 10.1007/s00449-018-02064-8 URL
[13]	黄蒙蒙, 何剑波, 石会玲, 等. Chlorella vulgar HDA04生长条件的初步确立[J]. 中国农学通报, 2021, 37(12):79-85.
[14]	LIN Y, GE J, LING H, et al. Isolation of a novel strain of Monoraphidium sp. and characterization of its potential for α-linolenic acid and biodiesel production[J]. Bioresource technology, 2018, 267:466-472. doi: 10.1016/j.biortech.2018.07.081 URL
[15]	LIN Y, GE J, ZHANG Y, et al. Monoraphidium sp. HDMA-20 is a new potential source of α-linolenic acid and eicosatetraenoic acid[J]. Lipids in health and disease, 2019, 18(1):56. doi: 10.1186/s12944-019-0996-5 URL
[16]	GORMAN D S, LEVINE R P. Cytochrome f and plastocyanin: their sequence in the photosynthetic electron transport chain of Chlamydomonas reinhardi[J]. Proc natl acad sci U.S.A, 1965, 54(6):1665-1669. doi: 10.1073/pnas.54.6.1665 URL
[17]	GREENSPAN P, MAYER E P, FOWLER S D. Nile red: a selective fluorescent stain for intracellular lipid droplets[J]. Journal of cell biology, 1985, 100(3):965-73.
[18]	ILAVARASI A, MUBARAKALI D, PRAVEENKUMAR R, et al. Optimization of various growth media to freshwater microalgae for biomass production[J]. Biotechnology, 2011, 10(6):540-545.
[19]	GEORGE B, PANCHA I, DESAI C, et al. Effects of different media composition, light intensity and photoperiod on morphology and physiology of freshwater microalgae Ankistrodesmus falcatus -a potential strain for bio-fuel production[J]. Bioresource technology, 2014, 171:367-374. doi: 10.1016/j.biortech.2014.08.086 URL
[20]	VES-URAI P, KROBTHONG S, THONGSUK K, et al. Comparative secretome analysis between salinity-tolerant and control Chlamydomonas reinhardtii strains[J]. Planta, 2021, 253(3):1-17. doi: 10.1007/s00425-020-03501-3 URL
[21]	YEE W. Feasibility of various carbon sources and plant materials in enhancing the growth and biomass productivity of the freshwater microalgae Monoraphidium griffithii NS16[J]. Bioresour Technol, 2015, 196:1-8. doi: 10.1016/j.biortech.2015.07.033 URL
[22]	WU L F, CHEN P C, LEE C M. The effects of nitrogen sources and temperature on cell growth and lipid accumulation of microalgae[J]. International biodeterioration & biodegradation, 2013, 85:506-510.
[23]	SALBITANI G and CARFAGNA S. Ammonium utilization in microalgae: a sustainable method for wastewater treatment[J]. Sustainability, 2021, 13(2):956. doi: 10.3390/su13020956 URL
[24]	LIN W, LI P, LIAO Z, et al. Detoxification of ammonium to Nannochloropsis oculata and enhancement of lipid production by mixotrophic growth with acetate[J]. Bioresource technology, 2017, 227:404. doi: 10.1016/j.biortech.2016.12.093 URL
[25]	代亮亮, 李莉杰, 郭亮亮, 等. 铵态氮和浮游植物对穗花狐尾藻的生理影响[J]. 生态学杂志, 2019, 38(9):2727-2733.
[26]	高怀骏, 陆庆楠, 庄铁钢, 等. 氮磷浓度变化下粉绿狐尾藻的生长规律研究[J]. 中国农学通报, 2021, 37(3):66-72.

产油单针藻的最适培养基筛选及产油特性提升

Lipid-producing Monoraphidium sp.: The Best Medium Screening and Lipid-producing Property Enhancement

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 8

参考文献 26

相关文章 15

编辑推荐

Metrics

本文评价

关于本刊

作者中心

审者中心

在线期刊

联系我们

[1]	董文彩, 刘宪斌, 李红梅, 赵双梅, 包金美, 沈健萍, 梁芳, 鲁美. 不同水平供钙量对木本观赏植物生长发育的影响[J]. 中国农学通报, 2022, 38(8): 42-50.
[2]	赵双梅, 刘宪斌, 李红梅, 董文彩, 沈健萍, 包金美, 梁芳, 鲁美. 云南哀牢山湿性常绿阔叶林土壤碳分布特征[J]. 中国农学通报, 2022, 38(8): 88-95.
[3]	段青, 蔡晶晶, 杜文文, 杨楠, 王祥宁, 贾文杰, 马璐琳. 大丽花花粉活力及离体萌发研究[J]. 中国农学通报, 2022, 38(7): 52-61.
[4]	李雪枫, 王坚, 叶晓园, 张秀婷, 王丽学. 苦瓜植株水浸提液对水稻种子萌发和秧苗生长的影响[J]. 中国农学通报, 2022, 38(6): 1-7.
[5]	李露, 包欣怡, 陈智勇. 非洲紫罗兰离体快繁体系的建立[J]. 中国农学通报, 2022, 38(6): 58-62.
[6]	李佳欢, 王晓慧, 姜明国, 宋福强, 常伟. 基于响应面法优化印度梨形孢培养基[J]. 中国农学通报, 2022, 38(3): 102-109.
[7]	吴文. 切根对乌桕容器苗质量的影响[J]. 中国农学通报, 2022, 38(3): 25-29.
[8]	梁培鑫, 唐榕, 郭晨荔, 郭睿, 何皇成, 王腾飞, 刘建国. 油莎豆对自然盐碱胁迫的生长及生理响应[J]. 中国农学通报, 2022, 38(26): 1-8.
[9]	连晓倩, 陶长铸, 郭昊澜, 李娜娜, 曹越, 吴鹏飞. 光照对芦苇生物生产力及光合能力的影响[J]. 中国农学通报, 2022, 38(20): 47-52.
[10]	马想, 蔡永立, 梁晶. 有机改良材料对滨海吹填土肥力质量的影响[J]. 中国农学通报, 2022, 38(19): 120-127.
[11]	白艳荣, 蒋亚莲, 王进英. 腊花组培快繁体系研究[J]. 中国农学通报, 2022, 38(16): 46-50.
[12]	杨晓燕, 叶伟伟, 魏善强, 张龙, 黄俊. 一株巨大芽孢杆菌发酵培养基的优化及解磷效果研究[J]. 中国农学通报, 2022, 38(15): 105-112.
[13]	王艳, 肖洋. 不同施肥处理对谷稗生物量和品质的影响[J]. 中国农学通报, 2022, 38(12): 20-25.
[14]	明佳佳, 向极钎, 康宇, 黄林, 陈永波, 瞿勇, 胡百顺, 殷红清. 不同钝化剂对恩施Cd污染土壤的钝化效果研究[J]. 中国农学通报, 2022, 38(1): 93-99.
[15]	朱海云, 马瑜, 柯杨, 李勃. 抗猕猴桃细菌性溃疡病蜡样芽孢杆菌MA23培养基及发酵条件优化[J]. 中国农学通报, 2021, 37(7): 112-118.