Exploration, Identification and Phosphorus-solubilizing Condition Optimization of a Multifunctional Strain

doi:10.13560/j.cnki.biotech.bull.1985.2022-1450

Abstract

Abstract:

In order to explore multifunctional microorganism resources to achieve the effective utilization of agricultural waste, some efficient phosphate-solubilizing bacteria were isolated from the compost of mushroom chaff and chicken manure, and their phosphate-solubilizing abilities were subsequently optimized. Efficient phosphorus-solubilizing bacteria were screened from the compost by inorganic phosphorus selective medium. The selected strain was identified through the combination of morphological observation, physiological and biochemical test and molecular biology identification. Single factor and response surface experiments were used to optimize the inorganic phosphorus-solubilizing conditions of this strain. In addition, the compost potential of this strain was evaluated by determining the ability of releasing organophosphorus and potassium, and indole acetic acid production. An efficient phosphate-solubilizing strain Pb1 was isolated and identified as belonging to Bacillus smithii strain. The highest phosphorus-solubilizing capacity of strain B. smithii Pb1 was achieved when the culture condition was kept as: glucose 15.0 g /L, ammonium sulfate 3 g/L, tricalcium phosphate 7.0 g/L, inorganic salt 0.48 g/L, temperature 55.0℃, rotating speed 225 r/min, initial pH 7.0, loaded liquid 60.0 mL/250 mL, and inoculum volume 3.0%（V/V）. Further response surface optimization showed that under the conditions of tricalcium phosphate 6.4 g/L, rotating speed 207 r/min and loaded liquid 65.0 mL/250 mL, the solubilized phosphorus content in the fermentation broth of strain Pb1 cultured for 3 dreached 534.68 mg/L, which was 1.58 times higher than that before optimization. Moreover, B. smithii Pb1 was found to be capable of detoxifying organophosphates, detoxifying potassium and producing indole-3-acetic acid with the content of 238.99 mg/L, 81.06 mg/L and 23.26 mg/L, respectively. B. smithii Pb1 demonstrated the ability of releasing inorganic phosphorus, organophosphates and potassium, and producing indoleacetic acid at high temperature（55℃）, laying the foundation for the development of B. smithii Pb1 as an aerobic composting bacterial agent.

Key words: multifunctional strain, screening, identification, characteristics of phosphorus solubilizing, response surface optimization

XIE Dong, WANG Liu-wei, LI Ning-jian, LI Ze-lin, XU Zi-hang, ZHANG Qing-hua. Exploration, Identification and Phosphorus-solubilizing Condition Optimization of a Multifunctional Strain[J]. Biotechnology Bulletin, 2023, 39(7): 241-253.

Figures/Tables 14

References 35

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因素Factors	水平Level
因素Factors	-1	+1
A: 葡萄糖浓度Glucose concentration/（g·L^-1）	15.0	20.0
B: 磷酸三钙浓度Concentration of tricalcium phosphate/（g·L^-1）	5.0	7.0
C: 硫酸铵浓度Ammonium sulfate concentration/（g·L^-1）	1.0	2.0
D: 无机盐浓度Concentration of inorganic salts/（g·L^-1）	0.48	0.96
E: 初始pH Initial pH	6.0	7.0
F: 温度Temperature/℃	50.0	55.0
G: 接种量Inoculum quantity/%	2.0	3.0
H: 装液量Loaded liquid/mL	60.0	80.0
J: 转速Rotating speed /（r·min^-1）	200	225

因素Factors	水平Level
因素Factors	-1	+1
A: 葡萄糖浓度Glucose concentration/（g·L^-1）	15.0	20.0
B: 磷酸三钙浓度Concentration of tricalcium phosphate/（g·L^-1）	5.0	7.0
C: 硫酸铵浓度Ammonium sulfate concentration/（g·L^-1）	1.0	2.0
D: 无机盐浓度Concentration of inorganic salts/（g·L^-1）	0.48	0.96
E: 初始pH Initial pH	6.0	7.0
F: 温度Temperature/℃	50.0	55.0
G: 接种量Inoculum quantity/%	2.0	3.0
H: 装液量Loaded liquid/mL	60.0	80.0
J: 转速Rotating speed /（r·min^-1）	200	225

因素Factor	水平Level
因素Factor	-1	0	+1
B: 磷酸三钙浓度 Concentration of tricalcium phosphate/（g·L^-1）	5.5	6.5	7.5
H: 装液量Loaded liquid/mL	55.0	60.0	75.0
J: 转速Rotating speed/（r·min^-1）	195	207	219

因素Factor	水平Level
因素Factor	-1	0	+1
B: 磷酸三钙浓度 Concentration of tricalcium phosphate/（g·L^-1）	5.5	6.5	7.5
H: 装液量Loaded liquid/mL	55.0	60.0	75.0
J: 转速Rotating speed/（r·min^-1）	195	207	219

指标 Index	结果 Result
接触酶试验 Catalase test	+
淀粉水解 Amylohydrolysis	+
明胶液化 Gelatin liquefaction	-
尿素酶反应 Urease reaction	+
甲基红（M.R）试验 Methyl red test	-
柠檬酸盐利用 Utilization of citrate	-
吲哚试验 Indole test	+
葡萄糖发酵 Fermentation of glucose	+
蔗糖发酵反应 Sucrose fermentation reaction	+