不同氮源及氮浓度对海水驯化藻株Asterarcys sp.生长及生化组成的影响

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

摘要/Abstract

摘要：

以经海水驯化后的藻株Asterarcys sp. SCSIO-44020为材料,以30‰海水改良ZSNT为基础培养基,在柱式光生物反应器中分别加入3种不同类型氮源（硝酸钠、尿素及碳酸氢铵）和3组氮浓度（3 mmol/L、6 mmol/L及18 mmol/L）,研究不同氮源及氮浓度对Asterarcys sp.生长及生化组成的影响。结果显示,Asterarcys sp.生物质浓度均随着氮浓度的升高而增加,在氮浓度18 mmol/L条件下,硝酸钠组获得最大生物质浓度（7.78 g/L）,其次为尿素（6.61 g/L）,最低为碳酸氢铵（5.33 g/L）。低氮胁迫有利于藻细胞油脂和总糖的积累,最大油脂含量和多糖含量分别达到46.78% DW（碳酸氢铵,3.0 mmol/L）和29.70% DW（尿素,3.0 mmol/L）。该藻中性脂比例大于82.58%,在3种氮源随着氮浓度的降低而升高;脂肪酸主要包括C16:0（棕榈酸）、C16:1（棕榈油酸）、C18:0（硬脂酸）、C18:1（油酸）、C18:2（亚油酸）和C18:3（亚麻酸）,其中油酸含量最高（占总脂含量38.75%）;高氮有利藻细胞蛋白质积累,在尿素为氮源18 mmol/L条件下的蛋白质含量最高（17.61% DW）;在以硝酸钠为氮源18 mmol/L条件下,该藻总脂、总糖、总蛋白质及总类胡萝卜素均获得最大产量,分别为2.79 g/L、1.71 g/L、1.22 g/L和30.29 mg/L。总之,本研究显示Asterarcys sp. SCSIO-44020在海水中生长良好,初步确定高浓度（18 mmol/L）硝酸钠为其小型柱式光生物反应器培养较佳的氮源条件。

关键词: Asterarcys sp., 氮源, 生化组成, 海水驯化

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

卫华宁, 王灵, 李涛, 王娜, 吴华莲, 向文洲. 不同氮源及氮浓度对海水驯化藻株Asterarcys sp.生长及生化组成的影响[J]. 生物技术通报, 2021, 37(10): 34-44.

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.

图/表 9

图1 不同氮源及氮浓度对Asterarcys sp. SCSIO-44020生长的影响 A:硝酸钠;B:尿素;C:碳酸氢铵

Fig.1 Effects of different nitrogen sources and concentra-tions on the growth of Asterarcys sp. SCSIO-44020 A:Sodium nitrate. B:Urea. C:Ammonium bicarbonate

图2 不同氮源及氮浓度对Asterarcys sp. SCSIO-44020总脂含量的影响

Fig. 2 Effects of different nitrogen sources and concentra-tions on total lipid content of Asterarcys sp. SCSIO-44020

图3 不同氮源及氮浓度对Asterarcys sp. SCSIO-44020总脂组分的影响 a: 3.0 mmol/L;b:6.0 mmol/L;c:18.0 mmol/L;PLs:磷脂;GLs:糖脂;NLs:中性脂

Fig. 3 Effects of different nitrogen sources and concentra-tions on the lipid components of Asterarcys sp. SCSIO-44020 PLs:Phospholipid. GLs:Glycolipid. NLs:Neutral lipid

图4 不同氮源及氮浓度对Asterarcys sp. SCSIO-44020总糖含量的影响

Fig. 4 Effects of different nitrogen sources and concentra-tions on the carbohydrates content of Asterarcys sp. SCSIO-44020

图5 不同氮源及氮浓度对Asterarcys sp. SCSIO-44020粗蛋白含量的影响

Fig. 5 Effects of different nitrogen sources and concentra-tions on the protein content of Asterarcys sp. SCSIO-44020

表1 不同氮源及氮浓度对Asterarcys sp. SCSIO-44020脂肪酸组成的影响

Table 1 Effects of different nitrogen sources and concentrations on the fatty acid composition of Asterarcys sp. SCSIO-44020

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

图6 不同氮源及氮浓度对Asterarcys sp. SCSIO-44020总叶绿素含量的影响

Fig. 6 Effects of different nitrogen sources and concentra-tions on the chlorophyll content of Asterarcys sp. SCSIO-44020

图7 不同氮源及氮浓度对Asterarcys sp. SCSIO-44020类胡萝卜素含量的影响

Fig. 7 Effects of different nitrogen sources and concentra-tions on the carotenoids content of Asterarcys sp. SCSIO-44020

表2 不同氮源及氮浓度对Asterarcys sp. SCSIO-44020总脂、总糖、总蛋白质和总类胡萝卜素产量的影响

Table 2 Effects of different nitrogen sources and concentrations on the total lipid,carbohydrates,protein and carotenoids yields of Asterarcys sp.SCSIO-44020

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

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