Alkanes degradation characteristics of a moderately halophilic bacterium Salinicola sp. in a high salinity environment

doi:10.16085/j.issn.1000-6613.2018-1456

Abstract

Abstract:

A crude oil resistant strain X4 was isolated from oil contaminated soil near Qinghai oilfield. It was identified as Salinicola sp. and named as Salinicola sp. X4, since the 16Sr DNA sequencing homologie to Salinicola zeshunii strain N4^T (GenBank serial number EU056581) was up to 99%. The optimal temperature, NaCl concentration, pH, carbon source and nitrogen source of Salinicola sp. X4 was 30℃, 8%, 6.5, glycerol and NH₄Cl, respectively. The bacterium could produce bioemulsifier, and it had good emulsification ability toward some typical hydrocarbons such as octane, cetane, xylene. The CSH values of Salinicola sp. X4 could reach 60%. Using diesel oil (3g/L) as the sole carbon source in the inorganic salt medium containing 5% NaCl, the Salinicola sp. X4 could degrade 55% of the total diesel for five days. GC-MS analysis showed that the strain preferred to degrade medium-chain length hydrocarbons firstly; the average degradation rate of C₇―C₁₃, C₁₄―C₂₀, C₂₁―C₃₁ was 64.1%, 52.3% and 26.8%, respectively. The ionic surfactant SDS and TTAB had strong toxicity to the cells: strain X4 wasn’t able to survive when the concentration reached 100mg/L and 400mg/L, respectively; under the concentration of 40mg/L, the degradation rate of diesel oil was reduced to 20%. While the tolerance concentration of the strain to the non-ionic surfactants――Twain80 and bio-surfactant――rhamnolipid reached 400mg/L; and rhamnolipid is a suitable compound surfactant for strain X4.

Key words: moderately halophilic bacterium, alkanes degradation, diesel oil, surfactant

摘要：

从青海油田附近被石油污染的土壤中分离得到一株可利用原油为唯一碳源的菌株，将其命名为X4菌株。经16SrDNA分析鉴定，该菌株与中度嗜盐菌Salinicola zeshunii strain N4^T（GenBank序列号为EU056581）同源性高达99%。X4菌株的最适温度为30℃，最适盐度为8%，最适pH为6.5，最佳碳源为甘油，最佳氮源为氯化铵。该菌可产生生物乳化剂，具有较强的细胞疏水性，对正辛烷、十六烷、二甲苯等典型烃类物质具有良好的乳化能力，细胞CSH值达到60%以上。在含5%盐度的无机盐培养基中，以3g/L的柴油为唯一碳源，采用GC-MS定量分析X4菌株的烃降解特征，结果表明菌株X4培养5天后柴油的总降解率达56%，菌株X4优先降解中长链烃类；C₇～C₁₃烃类的平均降解率为64.1%，C₁₄～C₂₀烃类的平均降解率为52.3%，C₂₁～C₃₁烃类的平均降解率约26.8%。离子型表面活性剂TTAB和SDS对X4菌株生长具有较强的毒性：在浓度达到100mg/L和400mg/L时能完全抑制菌体生长；在40mg/L的浓度下，使得菌株对柴油的降解率降低到20%。而X4菌株对非离子型表活剂——吐温80和生物表面活性剂——鼠李糖脂的耐受浓度均可达400mg/L。鼠李糖脂是嗜盐菌X4菌株的合适复配表活剂。

关键词: 中度嗜盐菌, 降解, 柴油, 表面活性剂

CLC Number:

Jiahui LIN, Dan WANG, Shuang LI. Alkanes degradation characteristics of a moderately halophilic bacterium Salinicola sp. in a high salinity environment[J]. Chemical Industry and Engineering Progress, 2019, 38(04): 1894-1902.

林佳辉, 王丹, 李霜. 一株中度嗜盐菌Salinicola sp.在高盐环境中的烷烃降解特性[J]. 化工进展, 2019, 38(04): 1894-1902.

Figures/Tables 18

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菌株	细胞形态	细胞大小/μm	耐盐范围/g·L^-1	最佳盐度/g·L^-1	pH耐受范围	温度耐受范围/℃
X4	短杆状	(1.5～2.0)×(0.7～1.0)	1~250	50~100	4.5～9.5	10～45
SMB35	杆状	(1.1～1.4)×(0.5～0.6)	5~300	30~100	5.0～9.0	15～40
DSM 9502	杆状	(2.0～2.6)×(0.4～0.6)	0~180	90	7.0～11.0	10～45
AL12	杆状	(1.2～1.4)×(0.7～0.8)	30~150	80	6.0～10.0	15～50
TD01	短杆状	(1.5～2.0)×(0.8～1.2)	10~200	50~60	5.0～11.0	0～45

菌株	细胞形态	细胞大小/μm	耐盐范围/g·L^-1	最佳盐度/g·L^-1	pH耐受范围	温度耐受范围/℃
X4	短杆状	(1.5～2.0)×(0.7～1.0)	1~250	50~100	4.5～9.5	10～45
SMB35	杆状	(1.1～1.4)×(0.5～0.6)	5~300	30~100	5.0～9.0	15～40
DSM 9502	杆状	(2.0～2.6)×(0.4～0.6)	0~180	90	7.0～11.0	10～45
AL12	杆状	(1.2～1.4)×(0.7～0.8)	30~150	80	6.0～10.0	15～50
TD01	短杆状	(1.5～2.0)×(0.8～1.2)	10~200	50~60	5.0～11.0	0～45

菌株	盐度 /g·L^-1	种类	浓度	时间 /天	降解率/%
假单胞菌（Pseudomonas sp.）^[28]	30	原油	0.5g·L^-1	5	68
假单胞菌MZ01 ^[29]	<10	原油	0.2g·L^-1	9	55
红球菌Rhodococcus erythropolis）^[30]	<10	原油	2g·L^-1	8	58
芽孢杆菌（Bacillus subtils）^[31]	50	原油	2g·L^-1	7	60
中度嗜盐菌^[32]	50	多环芳烃	0.05g·L^-1		37
解脂耶罗维亚酵母^[33]	<10	柴油	1mL·L^-1	9	78
中度嗜盐菌（Salinicola sp. W1）^[15]	24	柴油	0.1g·L^-1		95
假单胞菌（Pseudomonous sp.）^[34] 芽孢杆菌（Bacillus sp.）^[34]	<10	柴油	10g·L^-1	7	18 31
中度嗜盐菌（Salinicola sp. X4）	50	柴油	3g·L^-1	5	56

菌株	盐度 /g·L^-1	种类	浓度	时间 /天	降解率/%
假单胞菌（Pseudomonas sp.）^[28]	30	原油	0.5g·L^-1	5	68
假单胞菌MZ01 ^[29]	<10	原油	0.2g·L^-1	9	55
红球菌Rhodococcus erythropolis）^[30]	<10	原油	2g·L^-1	8	58
芽孢杆菌（Bacillus subtils）^[31]	50	原油	2g·L^-1	7	60
中度嗜盐菌^[32]	50	多环芳烃	0.05g·L^-1		37
解脂耶罗维亚酵母^[33]	<10	柴油	1mL·L^-1	9	78
中度嗜盐菌（Salinicola sp. W1）^[15]	24	柴油	0.1g·L^-1		95
假单胞菌（Pseudomonous sp.）^[34] 芽孢杆菌（Bacillus sp.）^[34]	<10	柴油	10g·L^-1	7	18 31
中度嗜盐菌（Salinicola sp. X4）	50	柴油	3g·L^-1	5	56

时间/min	组分	降解率/%	平均降解率/%
5.89	C₇	55.5	64.1
7.20	C₁₀	58.5
7.72	C₁₁	82.8
8.63	C₁₃	74.5
8.79	C₁₂	69.2
10.01	C₁₄	71.2	52.3
11.69	C₁₄	72.3
14.35	C₁₆	53.6
17.31	C₁₇	51.0
20.45	C₁₈	44.2
23.62	C₂₀	33.6
26.79	C₂₁	40.3	26.8
27.51	C₂₃	13.1
29.87	C₂₉	38.2
33.91	C₃₁	15.7