Effects of Different Nitrogen Sources and Concentrations on the Growth and Biochemical Composition of Asterarcys sp. Accimated by Seawater

doi:10.13560/j.cnki.biotech.bull.1985.2021-0139

Abstract

Abstract:

In order to explore the effects of different nitrogen sources and concentrations on the growth and biochemical composition of marine seawater-acclimated algae Asterarcys sp. SCSIO-44020,three nitrogen concentrations（3,6 and 18 mmol/L）of three nitrogen sources（sodium nitrate,urea and ammonium bicarbonate）were added to 30‰ seawater ZSNT based medium. Results showed that the biomass concentration of Asterarcys sp. increased with the increase of nitrogen concentration. At 18 mmol/L,the highest biomass concentration（7.78 g/L）was obtained with sodium nitrate as nitrogen source,followed by urea（6.61 g/L）,and the lowest was obtained with ammonium bicarbonate（5.33 g/L）. Nitrogen limitation was beneficial to the accumulation of lipid and carbohydrates of algae cell,and the maximum lipid content and carbohydrates content reached 46.78% DW（ammonium bicarbonate,3.0 mmol/L）and 29.70% DW（urea,3.0 mmol/L）respectively. The results also showed that the ratio of neutral lipid was very high（≥82.58%）,and it increased with the decrease of nitrogen concentration under three nitrogen sources. The fatty acid composition of the Asterarcys sp. mainly included C16:0（palmitic acid）,C16:1（palmitoleic acid）,C18:0（stearic acid）,C18:1（oleic acid）,C18:2（linoleic acid）and C18:3（α-linolenic acid）,in which the content of oleic acid was the highest（38.75% of total fatty acid）. The high nitrogen was beneficial to the protein accumulation,and the protein content of Asterarcys sp. was the highest（17.61% DW）under the condition of 18 mmol/L urea as nitrogen source with sufficient nitrogen. The maximum production of total lipid,total carbohydrates,total protein and total carotenoids in this microalga was obtained at 18 mmol/L using sodium nitrate,which was 2.79,1.71,1.22,and 30.29 mg/L,respectively. Therefore,Asterarcys sp. SCSIO-44020 grew well in seawater,and high concentration（18 mmol/L）of sodium nitrate was initially determined as the optimal nitrogen source for its cultivation in column glass photobioreactor.

Key words: Asterarcys sp., nitrogen source, biochemical composition, seawater acclimation

WEI Hua-ning, WANG Ling, LI Tao, WANG Na, WU Hua-lian, XIANG Wen-zhou. Effects of Different Nitrogen Sources and Concentrations on the Growth and Biochemical Composition of Asterarcys sp. Accimated by Seawater[J]. Biotechnology Bulletin, 2021, 37(10): 34-44.

Figures/Tables 9

References 31

[1]	梁晶晶, 蒋霞敏, 叶丽, 等. 氮、磷、铁对三角褐指藻诱变株生长、总脂及脂肪酸的影响[J]. 生态学杂志, 2016, 35(1):189-198.
	Liang JJ, Jiang XM, Ye L, et al. Effects of nitrogen, phosphorus and iron on the growth, total lipid content and fatty acid composition of Phaeodactylum tricornutum mutant strain[J]. Chinese Journal of Ecology, 2016, 35(1):189-198.
[2]	戴晨明, 高保燕, 苏敏, 等. 光强及氮浓度对丝状绿藻双星藻生长及生化组成的影响[J]. 微生物学通报, 2020, 47(1):172-181.
	Dai CM, Gao BY, Su M, et al. Effects of light intensity and nitrogen concentration on the growth and biochemical composition of filamentous green alga Zygnema sp.[J]. Microbiology China, 2020, 47(1):172-181.
[3]	Hegewald E, Wolf M, Keller A, et al. ITS2 sequence-structure phylogeny in the Scenedesmaceae with special reference to Coelastrum(Chlorophyta, Chlorophyceae), including the new genera Coma-siella and Pectinodesmus[J]. Phycologia, 2010, 49(4):325-335. doi: 10.2216/09-61.1 URL
[4]	Hong JW, Kim SA, Chang JW, et al. Isolation and description of a Korean microalga, Asterarcys quadricellulare KNUA020, and analysis of its biotechnological potential[J]. Korean Journal of Pesticide Science, 2012, 27(3):197-203.
[5]	Varshney P, Beardall J, Bhattacharya S, et al. Effect of elevated carbon dioxide and nitric oxide on the physiological responses of two green algae, Asterarcys quadricellulare and Chlorella sorokiniana[J]. Journal of Applied Phycology, 2020, 32(1):189-204. doi: 10.1007/s10811-019-01950-2
[6]	Li T, Yang F, Xu J, et al. Evaluating differences in growth, photosynthetic efficiency, and transcriptome of Asterarcys sp. SCS-1881 under autotrophic, mixotrophic, and heterotrophic culturing conditions[J]. Algal Research, 2020, 45:101753. doi: 10.1016/j.algal.2019.101753 URL
[7]	Syrett PJ, Leftley JW. Nitrate and urea assimilation by algae[J]. Perspectives in Experimental Biology, 2016, 2:221-234.
[8]	Alonso DL, Belarbi EH, Rodríguez-Ruiz J, et al. Acyl lipids of three microalgae[J]. Phytochemistry, 1998, 47(8):1473-1481. doi: 10.1016/S0031-9422(97)01080-7 URL
[9]	Gao B, Xia S, Lei X, Zhang C, et al. Combined effects of different nitrogen sources and levels and light intensities on growth and fatty acid and lipid production of oleaginous eustigmatophycean microalga Eustigmatos cf. polyphem[J]. Journal of Applied Phycology, 2018, 30(1):215-229. doi: 10.1007/s10811-017-1180-9 URL
[10]	Illman AM, Scragg AH, Shales SW, et al. Increase in Chlorella strains calorific values when grown in low nitrogen medium[J]. Enzyme and Microbial Technology, 2000, 27(8):631-635. pmid: 11024528
[11]	Arumugam M, Agarwal A, Arya MC, et al. Influence of nitrogen sources on biomass productivity of microalgae Scenedesmus bijugatus[J]. Bioresource Technology, 2013, 131:246-249. doi: 10.1016/j.biortech.2012.12.159 pmid: 23353039
[12]	吴桂秀, 黄罗冬, 高保燕, 等. 不同氮源及其浓度对标志链带藻合成淀粉和油脂的影响[J]. 微生物学报, 2016(7):1168-1177.
	Wu GX, Huang LD, Gao BY, et al. Effects of different nitrogen sources and concentrations on starch and lipid biosynjournal by Desmodesmus insignis[J]. Acta Microbiologica Sinica, 2016(7):1168-1177.
[13]	吴伯堂, 何汝洪, 彭云辉. 钝顶螺旋藻海水驯化的初步研究[J]. 海洋与湖沼, 1988, 19(2):197-200.
	Wu BT, He RH, Peng YH. A preliminary study of the effects of NaCl crude salt, and sea water on spirulina growth[J]. Oceanologia et Limnologia Sinica, 1988, 19(2):197-200.
[14]	李嘉颖, 李涛, 谭丽, 等. 盐度对一株淡水栅藻Scenedesmus sp. 生长及生化组成的影响[J]. 生物技术通报, 2017, 33(7):155-161.
	Li JY, Li T, Tan L, et al. Effects of salinity on the growth and biochemical properties of a freshwater algae Scenedesmus sp.[J]. Biotechnology Bulletin, 2017, 33(7):155-161.
[15]	Khozin-Goldberg I, Shrestha P, Cohen Z. Mobilization of arachidonyl moieties from triacylglycerols into chloroplastic lipids following recovery from nitrogen starvation of the microalga Parietochloris incisa[J]. BBA-Molecular and Cell Biology of Lipids, 2005, 1738(1-3):63-71.
[16]	Bigogno C, Khozin-Goldberg I, Boussiba S, et al. Lipid and fatty acid composition of the green oleaginous alga Parietochloris incisa, the richest plant source of arachidonic acid[J]. Phytochemistry, 2002, 60(5):497-503. pmid: 12052516
[17]	Scheiner D. Determination of ammonia and Kjeldahl nitrogen by indophenol method[J]. Water Research, 1976, 10(1):31-36. doi: 10.1016/0043-1354(76)90154-8 URL
[18]	Li T, Xu J, Gao B, et al. Morphology, growth, biochemical composition and photosynthetic performance of Chlorella vulgaris(Trebouxiophyceae)under low and high nitrogen supplies[J]. Algal Research, 2016, 16:481-491. doi: 10.1016/j.algal.2016.04.008 URL
[19]	Pruvost J, Vooren GV, Cogne G, et al. Investigation of biomass and lipids production with Neochloris oleoabundans in photobioreactor[J]. Bioresource Technology, 2009, 100(23):5988-5995. doi: 10.1016/j.biortech.2009.06.004 pmid: 19560349
[20]	许海, 陈丹, 陈洁, 等. 氮磷形态与浓度对铜绿微囊藻和斜生栅藻生长的影响[J]. 中国环境科学, 2019, 39(6):2560-2567.
	Xu H, Chen D, Chen J, et al. Effects of nitrogen and phosphorus forms and concentrations on the growth of Microcystis aeruginosa and Scenedesmus obliquus[J]. China Environmental Science, 2019, 39(6):2560-2567.
[21]	Muro-Pastor MI, Florencio FJ. Regulation of ammonium assimilation in cyanobacteria[J]. Plant Physiology & Biochemistry, 2003, 41(6-7):595-603.
[22]	胡章喜, 安民, 段舜山, 等. 不同氮源对布朗葡萄藻生长、总脂和总烃含量的影响[J]. 生态学报, 2009(6):3288-3294.
	Hu ZX, An M, Duan SS, et al. Effects of nitrogen sources on the growth, contents of total lipids and total hydrocarbons of Botryococcus braunii[J]. Acta Ecologica Sinica, 2009(6):3288-3294.
[23]	赵伟, 李涛, 吴华莲, 等. 不同氮源及氮浓度对耐高盐真眼点藻生长、脂类积累及脂肪酸组成的影响[J]. 生物技术通报, 2019, 35(6):62-68.
	Zhao W, Li T, Wu HL, et al. Growth, lipid accumulation, and fat composition of Eustigmatos sp. under different nitrogen source and concentration[J]. Biotechnology Bulletin, 2019, 35(6):62-68.
[24]	罗宁, 张森, 刘平怀. 氮、磷对热带海洋富油微藻Desmodesmus sp. WC08生长及油脂积累的影响[J]. 食品工业科技, 2016, 37(4):223-227.
	Luo N, Zhang S, Liu PH, et al. Effects of nitrogen and phosphorus on cell growth and lipid accumulation of tropic ocean microalgae strain Desmodesmus sp. WC08[J]. Science and Technology of Food Industry, 2016, 37(4):223-227.
[25]	Dai G, Qiu B, Forchhammer K. Ammonium tolerance in the cyanobacterium Synechocystis sp. strain PCC 6803 and the role of the psbA multigene family[J]. Plant Cell & Environment, 2013, 37(4):840-851.
[26]	吴琼芳, 张莹, 罗舒怀, 等. 氮限制对普通小球藻积累油脂过程中生化组成与光合生理的影响[J]. 植物科学学报, 2016, 34(2):280-288.
	Wu QF, Zhang Y, Luo SH, et al. Effects of nitrogen limitation on biochemical composition and photosynthetic physiology during lipid accumulation in Chlorella vulgaris Beijierineck[J]. Plant Science Journal, 2016, 34(2):280-288.
[27]	刘金丽, 王俊峰, 刘天中, 等. 缺氮条件对栅藻油脂积累与光合作用的影响[J]. 海洋科学, 2013(7):13-19.
	Liu JL, Wang JF, Liu TZ, et al. The effects of nitrogen starvation on lipid accumulation and photosynjournal of Scenedesmus dimorphus[J]. Marine Sciences, 2013(7):13-19.
[28]	赵萍. 三角褐指藻富油培养条件的优化及活性物质分析[D]. 烟台:鲁东大学, 2013.
	Zhao P. Optimizationn of culture conditions for oil production of Pheaodactylum tricornutum and analysis of active substances in the cells[D]. Yantai:Ludong University, 2013.
[29]	郝宗娣, 刘平怀, 杨勋, 等. 两株热带淡水微藻的海水驯化及总脂含量变化[J]. 广东农业科学, 2013(4):111-113.
	Hao ZD, Liu PH, Yang X, et al. Salinty acclimation of Coelastrum reticulatum & Scenedesums sp. and variation of lipid content[J]. Guangdong Agricultural Sciences, 2013(4):111-113.
[30]	向文洲, 李涛, 吴华莲, 等. 海水螺旋藻产业发展战略研究[J]. 广西科学, 2014, 21(6):573-579.
	Xiang WZ, Li T, Wu HL, et al. The strategic studies on developing industry of seawater spirulina with efforts[J]. Guangxi Sciences, 2014, 21(6):573-579.
[31]	Singh DP, Khattar JS, Rajput A, et al. High production of carotenoids by the green microalga Asterarcys quadricellulare PUMCC 5. 1. 1 under optimized culture conditions[J]. PLoS One, 2019, 14(9):0221930.

Nitrogen source	Concentration/（mmol·L^-1）	% TFA
Nitrogen source	Concentration/（mmol·L^-1）	C16:0	C16:1	C18:0	C18:1	C18:2	C18:3	Other
NaNO₃	3.0	26.94±0.22	1.58±0.04	3.52±0.25	37.64±0.08	10.94±0.15	10.49±0.27	8.90±0.53
	6.0	25.37±0.14	1.92±0.01	3.96±0.03	37.53±0.12	12.55±0.09	9.17±0.03	9.50±0.08
	18.0	20.93±0.09	2.11±0.01	4.39±0.04	34.93±0.31	16.69±0.04	8.83±0.02	12.12±0.45
CH₄N₂O	3.0	26.87±0.11	1.86±0.01	3.36±0.03	37.01±0.11	10.78±0.04	10.87±0.17	9.23±0.04
	6.0	25.09±0.11	1.83±0.03	4.11±0.01	37.72±0.07	11.66±0.03	9.99±0.07	9.60±0.31
	18.0	21.66±0.01	2.36±0.02	4.07±0.01	33.47±0.13	17.79±0.18	9.90±0.16	10.75±0.50
NH₄HCO₃	3.0	27.34±0.04	2.05±0.05	2.95±0.05	36.13±0.15	11.83±0.02	10.65±0.06	9.04±0.05
	6.0	27.42±0.18	1.82±0.06	3.36±0.02	36.76±0.09	11.24±0.07	10.43±0.05	8.96±0.11
	18.0	26.20±0.28	1.68±0.02	4.51±0.07	38.75±0.15	11.65±0.13	8.61±0.23	8.60±0.15

Nitrogen source	Concentration/（mmol·L^-1）	% TFA
Nitrogen source	Concentration/（mmol·L^-1）	C16:0	C16:1	C18:0	C18:1	C18:2	C18:3	Other
NaNO₃	3.0	26.94±0.22	1.58±0.04	3.52±0.25	37.64±0.08	10.94±0.15	10.49±0.27	8.90±0.53
	6.0	25.37±0.14	1.92±0.01	3.96±0.03	37.53±0.12	12.55±0.09	9.17±0.03	9.50±0.08
	18.0	20.93±0.09	2.11±0.01	4.39±0.04	34.93±0.31	16.69±0.04	8.83±0.02	12.12±0.45
CH₄N₂O	3.0	26.87±0.11	1.86±0.01	3.36±0.03	37.01±0.11	10.78±0.04	10.87±0.17	9.23±0.04
	6.0	25.09±0.11	1.83±0.03	4.11±0.01	37.72±0.07	11.66±0.03	9.99±0.07	9.60±0.31
	18.0	21.66±0.01	2.36±0.02	4.07±0.01	33.47±0.13	17.79±0.18	9.90±0.16	10.75±0.50
NH₄HCO₃	3.0	27.34±0.04	2.05±0.05	2.95±0.05	36.13±0.15	11.83±0.02	10.65±0.06	9.04±0.05
	6.0	27.42±0.18	1.82±0.06	3.36±0.02	36.76±0.09	11.24±0.07	10.43±0.05	8.96±0.11
	18.0	26.20±0.28	1.68±0.02	4.51±0.07	38.75±0.15	11.65±0.13	8.61±0.23	8.60±0.15

Nitrogen source	Concentration /（mmol·L^-1）	Biomass/ （g·L^-1）	Lipid yield/ （g·L^-1）	Carbohydrates yield/ （g·L^-1）	Protein yield/ （g·L^-1）	Carotenoids yield/ （mg·L^-1）
NaNO₃	3.0	3.35	1.45	0.97	0.20	4.45
	6.0	5.02	2.21	1.23	0.41	8.85
	18.0	7.78	2.79	1.71	1.22	30.29
CH₄N₂O	3.0	3.63	1.49	1.08	0.15	4.81
	6.0	5.13	2.09	1.38	0.23	8.88
	18.0	6.61	2.14	1.42	1.17	25.27
NH₄HCO₃	3.0	1.95	0.91	0.45	0.17	3.41
	6.0	2.90	1.24	0.80	0.25	4.51
	18.0	5.33	2.45	1.28	0.62	11.22

Nitrogen source	Concentration /（mmol·L^-1）	Biomass/ （g·L^-1）	Lipid yield/ （g·L^-1）	Carbohydrates yield/ （g·L^-1）	Protein yield/ （g·L^-1）	Carotenoids yield/ （mg·L^-1）
NaNO₃	3.0	3.35	1.45	0.97	0.20	4.45
	6.0	5.02	2.21	1.23	0.41	8.85
	18.0	7.78	2.79	1.71	1.22	30.29
CH₄N₂O	3.0	3.63	1.49	1.08	0.15	4.81
	6.0	5.13	2.09	1.38	0.23	8.88
	18.0	6.61	2.14	1.42	1.17	25.27
NH₄HCO₃	3.0	1.95	0.91	0.45	0.17	3.41
	6.0	2.90	1.24	0.80	0.25	4.51
	18.0	5.33	2.45	1.28	0.62	11.22