Urease-driven preparation of calcium carbonate micro-nanoparticles with different polymorphs

doi:10.11949/0438-1157.20210378

Abstract

Abstract:

The urease produced by Sporosarcina pasteurii can hydrolyze urea, and the generated CO₂ reacts with calcium chloride to obtain calcium carbonate with different polymorphs by urease in different solutions. The mesoporous hollow micron pure spheroids formed by self-assembly of nanospheres were obtained by urease in the supernatant of fermentation broth. Calcite with 100% was obtained by bacterial urease. Infrared spectrum analysis showed that the elliptical calcium carbonate obtained by crude urease was affected by the hydroxyl groups. The effects of urease activity and reactant concentration on vaterite were explored. It was also found that the obtained pure vaterite is very stable within 7 days and is expected to be used as a drug carrier.

Key words: urease, enzyme, catalysis, vaterite, calcite, nanoparticles

摘要：

巴氏芽孢八叠球菌（Sporosarcina pasteurii）产生的脲酶可水解尿素，生成的CO₂与氯化钙反应获得碳酸钙晶体，不同形式的脲酶溶液对碳酸钙晶型有显著影响。用发酵液上清液中的脲酶催化，可以获得由纳米级球霰石自组装成的介孔空心微米纯球霰石；用菌体中的脲酶催化可获得100%的方解石。红外光谱分析表明，细胞破碎后的粗脲酶溶液获得的椭圆形碳酸钙受到富含羟基物质的影响。进一步探究了脲酶活性和反应物浓度对球霰石的影响，并发现获得的纯球霰石在7 d内非常稳定，有望作为药物载体使用。

关键词: 脲酶, 酶, 催化, 球霰石, 方解石, 纳米粒子

CLC Number:

Q 819

Hui ZHOU,Zhifeng TIAN,Xiaowei TANG,Zhilong XIU. Urease-driven preparation of calcium carbonate micro-nanoparticles with different polymorphs[J]. CIESC Journal, 2021, 72(10): 5319-5329.

周惠,田志锋,唐小微,修志龙. 脲酶驱动不同晶型碳酸钙微纳米颗粒的制备[J]. 化工学报, 2021, 72(10): 5319-5329.

Figures/Tables 13

Table 1 The results of urease activity in different solutions

脲酶溶液	脲酶活性/(U/L)	蛋白质含量/(g/L)	比活性/(U/g)
fermentation broth	3.12 ± 0.02	0.77 ± 0.03	4.06 ± 0.12
supernatant	1.24 ± 0.01	0.29 ± 0.04	4.28 ± 0.12
bacterial cell	3.75 ± 0.16	0.62 ± 0.02	6.05 ± 0.01
crude enzyme solution	5.75 ± 0.08	0.87 ± 0.03	6.63 ± 0.21

Fig.1 pH changes within 30 min during reaction catalyzed by urease in different solutions

Fig.2 SEM images and particle size distribution of CaCO3 precipitates prepared by urease in different solutions

Fig.3 XRD images of CaCO3 precipitation obtained by urease in different solutions

Table 2 Proportion of vaterite and calcite by urease in different solutions

脲酶溶液	球霰石	方解石
发酵液	88.10%	11.90%
上清液	100.00%	0
菌体	0	100.00%
粗酶液	53.26%	46.74%

Fig.4 Morphology of vaterite and CaCO3 crystal prepared by crude enzyme solution

Fig.5 Infrared spectra of CaCO3 precipitates

Fig.6 Effect of urease activity on vaterite

Fig.7 Effect of reactant concentration on vaterite

Fig.8 Average particle size of vaterite under different reactant concentrations at < 1 min and 30 min

Fig.9 SEM of calcite and vaterite at different time(a)—(d) were the crystal morphology of the control group at <1 min, 30 min, 7 d and 14 d, respectively; (e)—(h) were the crystal morphology of the pure vaterite group at <1 min, 30 min, 7 d and 14 d, respectively

Fig.10 XRD of control group and pure vaterite group at different time(a) control group, < 1 min; (b) control group, 30 min; (c) control group, 7 d; (d) control group, 14 d; (e) vaterite group, < 1 min; (f) vaterite group, 30 min; (g) vaterite group, 7 d; (h) vaterite group, 14 d

Table 3 Proportion of vaterite and calcite in control group and pure vaterite group at different time

晶体类型		<1 min	30 min	7 d	14 d
对照组	方解石	96.81%	100%	100%	100%
对照组	球霰石	3.19%	0	0	0
纯球霰石组	方解石	0	0	29.21%	100%
纯球霰石组	球霰石	100%	100%	70.78%	0

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