Cell growth and lipid accumulation of Monoraphidium sp. QLZ-3 in walnut shell extracts with carbon dioxide

doi:10.16085/j.issn.1000-6613.2019-1220

Abstract

Abstract:

The effect of CO₂ on the cell growth and lipid accumulation of Monoraphidium sp. QLZ-3 in walnut shell extracts (WSE) was studied. Experimental results showed that the biomass and lipid productivities of QLZ-3 were 196.85mg/(L·d) and 97.52mg/(L·d) respectively under the presence of 12% CO₂, which were enhanced by 1.33-fold and 1.57-fold compared with the control group, respectively. The exogenous CO₂ upregulated the expression level of rbcL that was cultivated in WSE, and thus resulted in an increase in the CO₂ fixation. In addition, the 12% CO₂ increased the utilization of polyphenol, and increased the activities of the ACCase and ME but decreased those of PEPC of QLZ-3 in WSE. The results indicated that exogenous CO₂ could enhance the biomass and lipid productivity and reduce the cost of microalgae cultivation in WSE, which provided a new theoretical basis for the utilization of walnut shell and the industrial production of microalgae.

Key words: waste treatment, carbon dioxide, bioreactors, biodiesel, biomass

摘要：

以单针藻Monoraphidium sp. QLZ-3为对象，研究了CO₂对微藻在核桃壳提取液（walnut shell extracts，WSE）中生长及油脂积累的影响。结果显示，在12%的CO₂条件下，微藻在WSE中的生物量产率及油脂产率达到196.85mg/(L·d)和97.52mg/(L·d)，分别是对照组的1.33倍和1.57倍。WSE培养下，外源CO₂上调了微藻中核酮糖1,5-二磷酸羧化酶基因（ribulose 1,5-bisphosphate carboxylase/oxygenase，rbcL）的表达量，从而促进了CO₂的固定。此外，12% 的CO₂提高了微藻对WSE中多酚的利用，同时上调了乙酰辅酶A羧化酶（acetyl coenzyme A carboxylase，ACCase）和苹果酸酶（malic enzyme，ME）活性，下调了磷酸烯醇式丙酮酸羧化酶（phosphoenolpyruvate carboxylase，PEPC）活性。研究表明，CO₂可以提高WSE中微藻的生物量产率和油脂产率，降低培养成本，为核桃壳的资源化利用及微藻的工业化生产提供了新的理论基础。

关键词: 废物处理, 二氧化碳, 生物反应器, 生物柴油, 生物质

CLC Number:

Hailiang XING,Xunzan DONG,Benyong HAN,Shuxiang GENG,Delu NING,Ting MA,Xuya YU. Cell growth and lipid accumulation of Monoraphidium sp. QLZ-3 in walnut shell extracts with carbon dioxide[J]. Chemical Industry and Engineering Progress, 2020, 39(4): 1575-1582.

邢海亮,董训赞,韩本勇,耿树香,宁德鲁,马婷,余旭亚. 二氧化碳联合核桃壳提取液促进单针藻Monoraphidium sp. QLZ-3的生长和油脂积累[J]. 化工进展, 2020, 39(4): 1575-1582.

Figures/Tables 10

References 37

1	董训赞, 赵永腾, 余旭亚. 褪黑素诱导糖蜜废醪液培养单针藻Monoraphidium sp. FXY-10产生物燃料[J]. 化工进展, 2019, 38(5): 2421-2428.
	DONG Xunzan, ZHAO Yongteng, YU Xuya. Research on the biofuels production of Monoraphidium sp. FXY-10 by combined melatonin and molasses wastewater[J]. Chemical Industry and Engineering Progress, 2019, 38(5): 2421-2428.
2	吕旭, 孙仁胜, 张红兵. 微藻规模培养技术研究进展[J]. 应用化工, 2019, 48(6): 1487-1490.
	LÜ Xu, SUN Rensheng, ZHANG Hongbing. Development of mass cultivation technology for microalgae[J]. Applied Chemical Industry, 2019, 48(6): 1487-1490.
3	HOH D, WATSON S, KAN E. Algal biofilm reactors for integrated wastewater treatment and biofuel production: a review[J]. Chemical Engineering Journal, 2016, 287: 466-473.
4	TAN C H, CHEN C-Y, SHOW P L, et al. Strategies for enhancing lipid production from indigenous microalgae isolates[J]. Journal of the Taiwan Institute of Chemical Engineers,2016, 63: 189–194.
5	郭沛, 马荣江, 余南阳, 等. 基于微藻培养的沼液处理相关耦合技术进展[J]. 化工进展, 2019, 38(2): 1027-1037.
	GUO Pei, MA Rongjiang, YU Nanyang, et al. Recent progress in coupling technologies of biogas slurry treatment based on microalgae cultivation[J]. Chemical Industry and Engineering Progress, 2019, 38(2): 1027-1037.
6	冯贞, 方晓璞. 核桃加工副产物综合利用途径[J]. 中国油脂, 2018, 43(9): 71-74.
	FENG Zhen, FANG Xiaopu. Comprehensive utilization ways of by-products from walnut processing[J]. China Oils and Fats, 2018, 43(9): 71-74.
7	张双杰, 邢宝林, 黄光许, 等. 核桃壳水热炭对六价铬的吸附特性[J]. 化工进展, 2016, 35(3)：950-956.
	ZHANG Shuangjie, XING Baolin, HUANG Guangxu, et al. A study on adsorption of Cr(Ⅵ) by hydrothermal carbon from walnut shell[J]. Chemical Industry and Engineering Progress, 2016, 35(3)：950-956.
8	郑渝川, 黄仲华, 黄伊嘉, 等. 3种核桃副产物多酚提取液的体外抗氧化活性分析[J]. 四川林业科技, 2019, 40(1): 15-19.
	ZHENG Yuchuan, HUANG Zhonghua, HUANG Yijia, et al. A study of antioxidant activity in vitro of three kinds of walnut by-product polyphenol extracts[J]. Journal of Sichuan Forestry Science and Technology, 2019, 40(1): 15-19.
9	OUYANG H, HOU K, PENG W, et al. Antioxidant and xanthine oxidase inhibitory activities of total polyphenols from onion[J]. Saudi Journal of Biological Sciences, 2018, 25: 1509-1513.
10	ZHAO Y, WANG H, HAN B, et al. Coupling of abiotic stresses and phytohormonesfor the production of lipids and high-value by-products by microalgae: a review[J]. Bioresource Technology, 2019, 274: 549-556.
11	CHENG J, XU J, LU H, et al. Generating cycle flow between dark and light zones with double paddlewheels to improve microalgal growth in a flatplate photo-bioreactor[J]. Bioresource Technology,2018, 261: 151–157.
12	卫晴, 郑凌凌, 卢哲, 等. 两株绿藻响应CO₂浓度变化的生长和生理特性的研究[J]. 水生生物学报, 2018, 42 (1): 182-189.
	WEI Qing, ZHENG Lingling, LU Zhe, et al. The growth and physiological responses of two green algae to the change of CO₂ concentration[J]. Acta Hydrobiologica Sinica, 2018, 42 (1): 182-189.
13	ZHAO Y, LI D, DING K, et al. Production of biomass and lipids by the oleaginous microalgae Monoraphidium sp. QLY-1 through heterotrophic cultivation and photo-chemical modulator induction[J]. Bioresource Technology, 2016, 211: 669-676.
14	ZHAO Y, LI D, XU J W, et al. Melatonin enhances lipid production in, Monoraphidium sp. QLY-1 under nitrogen deficiency conditions via a multi-level mechanism[J]. Bioresource Technology, 2018, 259: 46-53.
15	KETHEESAN B, NIRMALAKHANDAN N. Feasibility of microalgal cultivation in a pilot-scale airlift-driven raceway reactor[J]. Bioresource Technology, 2012, 108: 196-202.
16	NIKESH K, AJAY B, RENU G. Shear rate and mass transfer coefficient in internal loop airlift reactors involving non-Newtonian fluids[J]. Chemical Engineering Research and Design, 2018, 136: 315-323.
17	PAZ-YÉPEZ C, PEINADO I, HEREDIA A, et al. Lipids digestibility and polyphenols release under in vitro digestion of dark, milk and white chocolate[J]. Journal of Functional Foods, 2019, 52: 196-203.
18	CHE R, HUANG L, XU J W, et al. Effect of fulvic acid induction on the physiology, metabolism, and lipid biosynthesis-related gene transcription of Monoraphidium sp. FXY-10[J]. Bioresource Technology, 2017, 227: 324-334.
19	CHE R, DING K, HUANG L, et al. Enhancing biomass and oil accumulation of Monoraphidium sp. FXY-10 by combined fulvic acid and two-step cultivation[J]. Journal of the Taiwan Institute of Chemical Engineers, 2016, 67: 161-165.
20	JIANG L, LUO S, FAN X, et al. Biomass and lipid production of marine microalgae using municipal wastewater and high concentration of CO₂[J]. Applied Energy, 2011, 88(10): 3336-3341.
21	MANORANJAN N, ANKUSH K, RAMKRISHNA S. Performance evaluation of microalgae for concomitant wastewater bioremediation, CO₂ biofixation and lipid biosynthesis for biodiesel application[J]. Algal Research, 2016, 16: 216-223.
22	郭琪, 郑凌凌, 沈伟, 等. 不同二氧化碳浓度培养对两株栅藻碳固定速率及油脂积累的影响[J]. 水生生物学报, 2016, 40(2): 414-418.
	GUO Qi, ZHENG Lingling, SHEN Wei, et al. Effect of different CO₂ concentration on CO₂ fixation and lipid accumulation of two strains of Scenedsmus sp.[J]. Acta Hydrobiologica Sinica, 2016, 40(2)：414-418.
23	李琴, 陈三凤. 适于猪场污水中快速生长富油微藻的筛选[J]. 农业生物技术学报, 2016, 24(7): 1083-1091.
	LI Qin, CHEN Sanfeng. Screening of oil-rich microalgae grown rapidly in swine farm wastewater[J]. Journal of Agricultural Biotechnology, 2016, 24(7): 1083-1091.
24	马红芳,李鑫,胡洪营, 等. 栅藻LX1在水产养殖废水中的生长、脱氮除磷和油脂积累特性[J]. 环境科学, 2012, 33(6): 1891-1896.
	MA Hongfang, LI Xin, HU Hongying, et al. Growth, removal of nitrogen and phosphorus, and lipid accumulation property of Scenedesmus sp. LX1 in aquaclture wastewater[J]. Environmental Sciences, 2012, 33(6): 1891-1896.
25	GHOSH A, KIRAN B. Carbon concentration in algae: reducing CO₂ from exhaust gas[J]. Trends Biotechnol. 2017, 35 (9): 806-880.
26	PEI H, JIANG L, HOU Q, et al. Toward facilitating microalgae cope with effluent from anaerobic digestion of kitchen waste: the art of agricultural phytohormones[J]. Biotechnology for Biofuels, 2017, 10: 76-93.
27	GUO Y, YUAN Z, XU J, et al. Metabolic acclimation mechanism in microalgae developed for CO₂ capture from industrial flue gas[J]. Algal Research, 2017, 26: 225-233.
28	LU Y, DU Y, QIN X, et al. Comprehensive evaluation of effective polyphenols in apple leaves and their combinatory antioxidant and neuroprotective activities[J]. Industrial Crops and Products, 2019, 129: 242-252.
29	LI D, ZHAO Y, DING W, et al. A strategy for promoting lipid production in green microalgae, Monoraphidium, sp. QLY-1 by combined melatonin and photoinduction[J]. Bioresource Technology, 2017, 235:104-112.
30	梅连阔, 魏强强, 张惠斌, 等.乙酰辅酶A羧化酶抑制剂的研究进展[J]. 中国药科大学学报, 2019, 50(3):253-264.
	MEI Liankuo, WEI Qiangqiang, ZHANG Huibin, et al. Research progress in acetyl-CoA carboxylase inhibitors[J]. Journal of China Pharmaceutical University, 2019, 50(3):253-264.
31	程万, 林辉, 赵宇华, 等. 苹果酸酶调控微生物油脂积累的研究进展[J]. 科技通报, 2010, 26(6): 853-857.
	CHENG Wan, LIN Hui, ZHAO Yuhua, et al. Research progress on malic enzyme regulating the accumulation of microbial oils[J]. Bulletin of Science and Technology, 2010, 26(6): 853-857.
32	XUE J, NIU Y F, HUANG T, et al. Genetic improvement of the microalga Phaeodactylum tricornutum for boosting neutral lipid accumulation[J]. Metabolic Engineering, 2015, 27: 1-9.
33	张金飞, 李霞, 何亚飞, 等. 外源葡萄糖增强高表达转玉米C₄型pepc水稻耐旱性的生理机制[J]. 作物学报, 2018(1):82-94.
	ZHANG Jinfei, LI Xia, HE Yafei, et al. Physiological mechanism on drought tolerance enhanced by exogenous glucose in C₄-pepc rice[J]. Acta Agronomica Sinica, 2018(1): 82-94.
34	王琳, 余旭亚, 赵鹏, 等. 微藻油脂生物合成与ACCase、PEPC相关性的研究进展[J]. 中国油脂, 2013, 38(2):56-60.
	WANG Lin, YU Xuya, ZHAO Peng, et al. Progress in relativity of lipid synthesis of microalgae with ACCase and PEPC[J]. China Oils and Fats, 2013, 38(2):56-60.
35	YANG J, PAN Y, BOWLER C, et al. Knockdown of phosphoenolpyruvate carboxykinase increases carbon flux to lipid synthesis in Phaeodactylum tricornutum[J]. Algal Reserach, 2016, 15: 50-58.
36	IKARAN Z, SUÁREZ-ALVAREZ S, URRETA I, et al. The effect of nitrogen limitation on the physiology and metabolism of Chlorella vulgaris var L3[J]. Algal Research, 2015, 10: 134-144.
37	YU X, ZHAO P, HE C, et al. Isolation of a novel strain of Monoraphidium sp. and characterization of its potential application as biodiesel feedstock[J]. Bioresource Technology, 2012, 121: 256-262.

引物	引物序列（5’-3’）	退火温度/℃
18S F	GGGAGTATGGTCGCAAGG	57
18S R	GACTATTTAGCAGGCTGAGGT
rbcL F	GTACCTGCCCAACCTCACCG	57
rbcL R	GCACCAGCATGGACACCACC

引物	引物序列（5’-3’）	退火温度/℃
18S F	GGGAGTATGGTCGCAAGG	57
18S R	GACTATTTAGCAGGCTGAGGT
rbcL F	GTACCTGCCCAACCTCACCG	57
rbcL R	GCACCAGCATGGACACCACC

脂肪酸	CO₂浓度
脂肪酸	对照组	4%	8%	12%	16%
C14∶0	0.28±0.0057	0.35±0.0099**	0.51±0.0021**	0.43±0.0014**	0.41±0.0057**
C16∶0	21.45±0.38	28.29±0.038**	30.55±0.049**	29.23±0.14**	29.30±0.049**
C16∶1	0.44±0.0085	0.14±0.0078**	0.17±0.0042**	0.20±0.0014**	0.28±0.013**
C18∶0	1.87±0.085	2.52±0.0078**	3.01±0.0042**	2.43±0.0035**	2.85±0.015**
C18∶1	24.06±0.089	29.05±0.012**	25.01±0.0057	24.24±0.0028	23.75±0.011
C18∶2	25.39±0.047	15.01±0.0049**	15.35±0.0085**	16.72±0.0071**	19.11±0.057**
C18∶3	22.76±0.12	19.18±0.0085**	20.33±0.0085*	21.76±0.0042	19.26±0.085**
C20∶0	0.16±0.014	0.22±0.0071**	0.24±0.0042**	0.21±0.0049**	0.24±0.0035**
C20∶1	1.44±0.022	1.89±0.0042**	1.47±0.0085	1.66±0.00071**	1.61±0.028**
C20∶3	0.026±0.0071	0.033±0.00071**	0.034±0.0035**	0.033±0.0014**	0.039±0.0028**
C22∶0	0.58±0.0028	0.9±0.028**	0.86±0.0078**	0.74±0.0049**	0.81±0.0085**
C22∶1	0.50±0.0092	0.75±0.0028**	0.69±0.0078**	0.66±0.0063**	0.63±0.0028**
C24∶0	0.35±0.0035	0.57±0.063**	0.59±0.011**	0.52±0.0063**	0.55±0.0028**
C24∶1	0.006±0.0014	0.028±0.0021**	0.029±0.00071**	0.008±0.0014**	0.026±0.0021**
SFA/%	24.69±0.47	32.85±0.048**	35.77±0.016**	33.57±0.023**	34.16±0.014**
MUFA/%	26.45±0.13	31.86±0.0078**	27.36±0.0028	26.77±0.013	26.29±0.021
PUFA/%	48.20±0.076	34.24±0.016**	35.71±0.011**	38.53±0.0099**	38.40±0.011**
DU/%	122.85±0.28	100.34±0.039**	98.77±0.020**	103.83±0.033**	103.10±0.044**
LCSF/%	4.81±0.069	6.80±0.076**	7.28±0.035**	6.51±0.0020**	6.91±00.024**
CFPP/℃	-16.32±0.0021	-16.3±0.0024	-16.25±0.0011	-16.27±0.0062	-16.26±0.0076

脂肪酸	CO₂浓度
脂肪酸	对照组	4%	8%	12%	16%
C14∶0	0.28±0.0057	0.35±0.0099**	0.51±0.0021**	0.43±0.0014**	0.41±0.0057**
C16∶0	21.45±0.38	28.29±0.038**	30.55±0.049**	29.23±0.14**	29.30±0.049**
C16∶1	0.44±0.0085	0.14±0.0078**	0.17±0.0042**	0.20±0.0014**	0.28±0.013**
C18∶0	1.87±0.085	2.52±0.0078**	3.01±0.0042**	2.43±0.0035**	2.85±0.015**
C18∶1	24.06±0.089	29.05±0.012**	25.01±0.0057	24.24±0.0028	23.75±0.011
C18∶2	25.39±0.047	15.01±0.0049**	15.35±0.0085**	16.72±0.0071**	19.11±0.057**
C18∶3	22.76±0.12	19.18±0.0085**	20.33±0.0085*	21.76±0.0042	19.26±0.085**
C20∶0	0.16±0.014	0.22±0.0071**	0.24±0.0042**	0.21±0.0049**	0.24±0.0035**
C20∶1	1.44±0.022	1.89±0.0042**	1.47±0.0085	1.66±0.00071**	1.61±0.028**
C20∶3	0.026±0.0071	0.033±0.00071**	0.034±0.0035**	0.033±0.0014**	0.039±0.0028**
C22∶0	0.58±0.0028	0.9±0.028**	0.86±0.0078**	0.74±0.0049**	0.81±0.0085**
C22∶1	0.50±0.0092	0.75±0.0028**	0.69±0.0078**	0.66±0.0063**	0.63±0.0028**
C24∶0	0.35±0.0035	0.57±0.063**	0.59±0.011**	0.52±0.0063**	0.55±0.0028**
C24∶1	0.006±0.0014	0.028±0.0021**	0.029±0.00071**	0.008±0.0014**	0.026±0.0021**
SFA/%	24.69±0.47	32.85±0.048**	35.77±0.016**	33.57±0.023**	34.16±0.014**
MUFA/%	26.45±0.13	31.86±0.0078**	27.36±0.0028	26.77±0.013	26.29±0.021
PUFA/%	48.20±0.076	34.24±0.016**	35.71±0.011**	38.53±0.0099**	38.40±0.011**
DU/%	122.85±0.28	100.34±0.039**	98.77±0.020**	103.83±0.033**	103.10±0.044**
LCSF/%	4.81±0.069	6.80±0.076**	7.28±0.035**	6.51±0.0020**	6.91±00.024**
CFPP/℃	-16.32±0.0021	-16.3±0.0024	-16.25±0.0011	-16.27±0.0062	-16.26±0.0076

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