中度嗜盐菌Halomonas salifodinae N35-6产四氢嘧啶的培养基优化

doi:10.6048/j.issn.1001-4330.2023.12.023

摘要/Abstract

摘要：

【目的】优化从新疆哈密七角井盐湖土壤中分离到一株四氢嘧啶产量较高的中度嗜盐菌N35-6发酵培养基。【方法】以四氢嘧啶产量为指标,采用响应面法优化培养基成分,利用HPLC检测四氢嘧啶产量。【结果】从七角井盐湖土壤中分离获得一株能够产四氢嘧啶的细菌,经16S rDNA 序列分析鉴定为Halomonas salifodinae,并命名为H.salifodinae N35-6,该菌株四氢嘧啶产量能够达到0.83 g/L。【结论】中度嗜盐菌Halomonas salifodinae N35-6具有高产四氢嘧啶的潜力。

关键词: 中度嗜盐菌; 四氢嘧啶; 优化

Abstract:

【Objective】A moderately halophilic bacterium Halomonas salifodinae N35-6 with high ectoine production was isolated from the soil of Qijaojing salt lake in Hami, Xinjiang, and its fermentation medium was optimized.【Methods】The medium composition for producing ectoine was optimized using response surface methodology and the yield of ectoine was determined by HPLC.【Results】A bacterium capable of producing ectoine was isolated from the soil of Qijiaojing Salt Lake and identified as Halomonas salifodinae by 16S rDNA sequence analysis. Production of ectoine was maximized to 0.83 g/L by optimizing the level of nutrients in the medium using response surface methodology.【Conclusion】The moderately halophilic bacterium Halomonas salifodinae N35-6 has the potential of high yield of ectoine.

Key words: moderately halophilic bacteria; ectoine; optimization

中图分类号:

S188

林青, 秦新政, 高雁, 曾军, 娄恺, 时红玲, 霍向东. 中度嗜盐菌Halomonas salifodinae N35-6产四氢嘧啶的培养基优化[J]. 新疆农业科学, 2023, 60(12): 3065-3071.

LIN Qing, Qin Xinzheng, GAO Yan, ZENG Jun, LOU Kai, SHI Hongling, HUO Xiangdong. Optimization of medium for ectoine production by Halomonas salifodinae N35-6, a moderately halophilic bacterium[J]. Xinjiang Agricultural Sciences, 2023, 60(12): 3065-3071.

图/表 5

表1 因子水平编码

Tab.1 Different variables with their levels

变量 Different variables	水平 Levels
变量 Different variables	-1	0	1
X₁:酵母粉(g/L)	5	10	15
X₂:L-谷氨酸钠(g/L)	10	25	40
X₃:NH₄Cl(g/L)	2	11	20
X₄:葡萄糖(g/L)	10	25	40
X₅:NaCl	50	100	150

图1 菌株Halomonas salifodinae N35-6系统进化树

Fig.1 The phylogenetic tree of Halomonas salifodinae N35-6

表2 响应面二次模型的方差

Tab.2 Analysis of variance of response surface quadratic model

来源 Source	自由度 Freedom	Seq SS	分布 Distribution	Adj SS	F 值 F value	P 值 P value
模型 Model	20	0.656 490	83.87%	0.656 490	6.50	0.000
线性 Linear	5	0.539 441	68.91%	0.539 441	21.36	0.000
yeast	1	0.403 437	51.54%	0.403 437	79.86	0.000
MSG	1	0.014 003	1.79%	0.014 003	2.77	0.108
NH₄Cl	1	0.047 560	6.08%	0.047 560	9.41	0.005
NaCl	1	0.067 947	8.68%	0.067 947	13.45	0.001
glucose	1	0.006 494	0.83%	0.006 494	1.29	0.268
平方Square	5	0.033 945	4.34%	0.033 945	1.34	0.279
yeast×yeast	1	0.012 647	1.62%	0.007 807	1.55	0.225
MSG×MSG	1	0.000 217	0.03%	0.000 008	0.00	0.968
NH₄Cl×NH₄Cl	1	0.003 460	0.44%	0.003 851	0.76	0.391
NaCl×NaCl	1	0.013 609	1.74%	0.009 018	1.79	0.194
glucose×glucose	1	0.004 011	0.51%	0.004 011	0.79	0.381
双因子交互作用 Two factor interaction	10	0.083 104	10.62%	0.083 104	1.65	0.151
yeast×MSG	1	0.014 762	1.89%	0.014 762	2.92	0.100
yeast×NH₄Cl	1	0.007 253	0.93%	0.007 253	1.44	0.242
yeast×NaCl	1	0.009 184	1.17%	0.009 184	1.82	0.190
yeast×glucose	1	0.011 413	1.46%	0.011413	2.26	0.145
MSG×NH₄Cl	1	0.016 087	2.06%	0.016 087	3.18	0.086
MSG×NaCl	1	0.008 130	1.04%	0.008 130	1.61	0.216
MSG×glucose	1	0.002 256	0.29%	0.002 256	0.45	0.510
NH₄Cl×NaCl	1	0.010 302	1.32%	0.010 302	2.04	0.166
NH₄Cl×glucose	1	0.000 136	0.02%	0.000 136	0.03	0.871
NaCl×glucose	1	0.003 580	0.46%	0.003 580	0.71	0.408
误差 Error	25	0.126 293	16.13%	0.126 293
失拟 Lacr of fit	20	0.111 613	14.26%	0.111 613	1.90	0.246
纯误差 Pure error	5	0.014 679	1.88%	0.014 679
合计 Total	45	0.782 783	100.00%
R²=0.838 7 $R_{a d j}^{2}$ =0.709 6

表2 响应面二次模型的方差

Tab.2 Analysis of variance of response surface quadratic model

来源 Source	自由度 Freedom	Seq SS	分布 Distribution	Adj SS	F 值 F value	P 值 P value
模型 Model	20	0.656 490	83.87%	0.656 490	6.50	0.000
线性 Linear	5	0.539 441	68.91%	0.539 441	21.36	0.000
yeast	1	0.403 437	51.54%	0.403 437	79.86	0.000
MSG	1	0.014 003	1.79%	0.014 003	2.77	0.108
NH₄Cl	1	0.047 560	6.08%	0.047 560	9.41	0.005
NaCl	1	0.067 947	8.68%	0.067 947	13.45	0.001
glucose	1	0.006 494	0.83%	0.006 494	1.29	0.268
平方Square	5	0.033 945	4.34%	0.033 945	1.34	0.279
yeast×yeast	1	0.012 647	1.62%	0.007 807	1.55	0.225
MSG×MSG	1	0.000 217	0.03%	0.000 008	0.00	0.968
NH₄Cl×NH₄Cl	1	0.003 460	0.44%	0.003 851	0.76	0.391
NaCl×NaCl	1	0.013 609	1.74%	0.009 018	1.79	0.194
glucose×glucose	1	0.004 011	0.51%	0.004 011	0.79	0.381
双因子交互作用 Two factor interaction	10	0.083 104	10.62%	0.083 104	1.65	0.151
yeast×MSG	1	0.014 762	1.89%	0.014 762	2.92	0.100
yeast×NH₄Cl	1	0.007 253	0.93%	0.007 253	1.44	0.242
yeast×NaCl	1	0.009 184	1.17%	0.009 184	1.82	0.190
yeast×glucose	1	0.011 413	1.46%	0.011413	2.26	0.145
MSG×NH₄Cl	1	0.016 087	2.06%	0.016 087	3.18	0.086
MSG×NaCl	1	0.008 130	1.04%	0.008 130	1.61	0.216
MSG×glucose	1	0.002 256	0.29%	0.002 256	0.45	0.510
NH₄Cl×NaCl	1	0.010 302	1.32%	0.010 302	2.04	0.166
NH₄Cl×glucose	1	0.000 136	0.02%	0.000 136	0.03	0.871
NaCl×glucose	1	0.003 580	0.46%	0.003 580	0.71	0.408
误差 Error	25	0.126 293	16.13%	0.126 293
失拟 Lacr of fit	20	0.111 613	14.26%	0.111 613	1.90	0.246
纯误差 Pure error	5	0.014 679	1.88%	0.014 679
合计 Total	45	0.782 783	100.00%
R²=0.838 7 $R_{a d j}^{2}$ =0.709 6

图2 因子影响四氢嘧啶产量的pareto 图

Fig.2 Pareto chart evaluates the variables affecting ectoine production

图3 Ectoine 产量影响因子交互作用曲面图

Fig.3 Interactions plots of variables affecting ectoine production

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