海洋产脲酶细菌的筛选及诱导形成碳酸钙矿物的特征

doi:10.19657/j.geoscience.1000-8527.2021.170

摘要/Abstract

摘要：

产脲酶细菌普遍存在于自然界,其对碳酸钙的矿化作用被广泛用于工程地质和环境等各项工程领域;本研究旨在从北戴河新河河口海洋沉积物中分离产脲酶细菌,探讨其诱导形成碳酸钙的特征,为地质与工程领域的应用提供良好的材料。从海洋沉积物样品中共分离出33株细菌,经筛选,其中有10株细菌具有产脲酶活性,16S rRNA基因序列分析表明:其中有3株细菌属于苍白杆菌属(Ochrobactrum sp.),而其他7株细菌属于赖氨酸芽孢杆菌属(Lysinibacillus sp.)。对其中两株有代表性的菌株人苍白杆菌(O. anthropic)CP57和赖氨酸芽孢杆菌(L. fusiformis) CP66诱导形成碳酸钙沉淀进行了实验,X射线晶体衍射(XRD)分析表明:CP57菌株诱导形成的沉淀由方解石组成,而CP66菌株诱导形成的沉淀则包含了方解石和球霰石两种晶相;扫描电子显微镜(SEM)观察显示:CP57形成的沉淀呈不规则形,而CP66形成的沉淀包含了不规则形和球形两种形态;进一步用透射电子显微镜(TEM)和选区电子衍射(SAED)对两种不同形态的碳酸钙沉淀进行了矿物相的分析,结果显示:不规则形碳酸钙沉淀为方解石,球形碳酸钙沉淀为球霰石;采用傅立叶变换红外光谱(FTIR)对细菌所形成的碳酸钙进行了表征,结果显示:CP57形成的沉淀分别在波数1 422 cm^-1、875 cm^-1和711 cm^-1处出现明显吸收峰,和方解石的FTIR图谱特征吻合;而CP66除出现以上方解石的吸收峰外,还在1 081 cm^-1和743 cm^-1处出现明显对应球霰石的特征吸收峰。研究结果为进一步揭示不同细菌诱导碳酸钙形成不同的形貌和晶相奠定了基础。

关键词: 产脲酶细菌, 生物矿化, 碳酸钙沉淀, 方解石, 球霰石

Abstract:

Urease-producing bacteria are widespread in nature, and the calcium carbonate biomineralization induced by these bacteria are widely used in various geological and engineering fields. This study aims to isolate the urease-producing bacteria from marine sediments, and to characterize the produced calcium carbonate crystals. The results may have potential applications in the geological and engineering fields. A total of 33 bacterial strains were isolated from the marine sediments sampled from Beidaihe (China), among which 10 bacterial strains were identified to produce urease. The 16S rRNA gene sequence analyses show that three strains belong to the Ochrobactrum genus, and the other seven strains belong to the Lysinibacillus genus. Two strains, O. anthropic CP57 and L. fusiformis CP66, were selected to characterize their calcium carbonate formations. The SEM, XRD, and SAED results show that the calcium carbonate formed by CP57 is irregular-shape calcite, whereas that formed by CP66 is irregular-shape or spherical calcite and vaterite. Further FTIR characterization of the bacterial calcium carbonate shows that the spectrum of CP57 at the wave numbers of 1,422 cm^-1, 711 cm^-1 and 875 cm^-1, correspond to the typical absorption peaks of calcite. However, exception appears in the FTIR spectrum of CP66, with two extra peaks at 1,081 cm^-1 and 743 cm^-1 clearly attributed to vaterite. Our results provide a foundation to reveal the mechanism of different morphologies and mineral phases formed by various bacteria.

Key words: urease-producing bacteria, biomineralization, calcium carbonate precipitation, calcite, vaterite

中图分类号:

P745

陈慧佳, 张慧卿, 冯莹, 魏士平. 海洋产脲酶细菌的筛选及诱导形成碳酸钙矿物的特征[J]. 现代地质, 2022, 36(01): 212-220.

CHEN Huijia, ZHANG Huiqing, FENG Ying, WEI Shiping. Screening of Marine Urease-producing Bacteria and Characterization of Their Produced Calcium Carbonate Minerals[J]. Geoscience, 2022, 36(01): 212-220.

图/表 6

图1 产脲酶细菌的筛选结果

Fig.1 Screening results of the urease-producing bacteria

表1 产脲酶细菌的16S rRNA基因序列比对结果

Table 1 Comparison of the 16S rRNA gene sequences of the urease-producing bacteria

菌株	属种	基因号	相似性/%	产脲酶活性
CP49	Ochrobactrum anthropi	KY770534	98.21	++
CP50	Lysinibacillus fusiformis	KU159195	98.67	++
CP51	Lysinibacillus fusiformis	KU159195	98.52	++
CP56	Ochrobactrum anthropi	MN176368	99.50	+++
CP57	Ochrobactrum anthropi	KY770534	98.09	+++
CP66	Lysinibacillus fusiformis	MN543803	98.91	++++
CP76	Lysinibacillus fusiformis	MW876146	99.37	+
CP77	Lysinibacillus fusiformis	KF208480	99.67	++
CP78	Lysinibacillus sphaericus	KU904281	99.77	+
CP79	Lysinibacillus sphaericus	KJ183074	98.95	+

图2 O. anthropic CP57和L. fusiformis CP66菌株诱导形成的碳酸钙XRD图谱

Fig.2 XRD patterns of calcium carbonate induced by O. anthropic CP57 and L. fusiformis CP66

图3 O. anthropi CP57和L. fusiformis CP66所形成碳酸钙沉淀的SEM图((a)—(c)CP57;(d)—(f)CP66)

Fig.3 SEM images of calcium carbonate precipitates formed by O. anthropi CP57 and L. fusiformis CP66((a)—(c)CP57;(d)—(f)CP66)

图4 O. anthropi CP57 (a)(c)和L. fusiformis CP66 (b)(d)所形成碳酸钙沉淀物的TEM (a)(b)及SAED (c)(d)图

Fig.4 TEM (a)(b) and SAED (c)(d) images of calcium carbonate precipitates formed by O. anthropi CP57 (a)(c) and L. fusiformis CP66 (b)(d)

图5 O. anthropic CP57和L. fusiformis CP66所形成的碳酸钙沉淀的FTIR图谱

Fig.5 FTIR spectra of calcium carbonate precipitations formed by O. anthropic CP57 and L. fusiformis CP66

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