Development of a semidefined growth medium for Pedobacter cryoconitis BG5 using statistical experimental design

M. Ong, Clarence M. Ongkudon, C. Wong
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引用次数: 2

Abstract

ABSTRACT Pedobacter cryoconitis BG5 are psychrophiles isolated from the cold environment and capable of proliferating and growing well at low temperature regime. Their cellular products have found a broad spectrum of applications, including in food, medicine, and bioremediation. Therefore, it is imperative to develop a high-cell density cultivation strategy coupled with optimized growth medium for P. cryoconitis BG5. To date, there has been no published report on the design and optimization of growth medium for P. cryoconitis, hence the objective of this research project. A preliminary screening of four commercially available media, namely tryptic soy broth, R2A, Luria Bertani broth, and nutrient broth, was conducted to formulate the basal medium. Based on the preliminary screening, tryptone, glucose, NaCl, and K2HPO4 along with three additional nutrients (yeast extract, MgSO4, and NH4Cl) were identified to form the basal medium which was further analyzed by Plackett–Burman experimental design. Central composite experimental design using response surface methodology was adopted to optimize tryptone, yeast extract, and NH4Cl concentrations in the formulated growth medium. Statistical data analysis showed a high regression factor of 0.84 with a predicted optimum optical (600 nm) cell density of 7.5 using 23.7 g/L of tryptone, 8.8 g/L of yeast extract, and 0.7 g/L of NH4Cl. The optimized medium for P. cryoconitis BG5 was tested, and the observed optical density was 7.8. The cost-effectiveness of the optimized medium was determined as 6.25 unit prices per gram of cell produced in a 250-ml Erlenmeyer flask.
采用统计实验设计开发一种半确定的冷冻结肠炎杆菌BG5生长培养基
低温嗜冷杆菌BG5是从寒冷环境中分离出来的嗜冷菌,能够在低温条件下增殖和生长。他们的细胞产品已经发现了广泛的应用,包括食品,医药和生物修复。因此,开发高细胞密度的培养策略和优化的生长培养基对冻结菌BG5的培养具有重要意义。到目前为止,还没有关于冷冻结膜假单胞菌生长培养基设计和优化的报道,因此本研究项目的目的是。初步筛选了四种市售培养基,即色氨酸大豆肉汤、R2A、Luria Bertani肉汤和营养肉汤,以制定基础培养基。在初步筛选的基础上,确定了色氨酸、葡萄糖、NaCl和K2HPO4以及另外三种营养物质(酵母提取物、MgSO4和NH4Cl)构成基础培养基,并通过Plackett-Burman实验设计进一步分析。采用响应面法进行中心复合实验设计,优化培养基中色氨酸、酵母提取物和NH4Cl的浓度。统计数据分析表明,色氨酸浓度为23.7 g/L,酵母浸膏浓度为8.8 g/L, NH4Cl浓度为0.7 g/L,预测最佳光学(600 nm)细胞密度为7.5,回归系数为0.84。对优化后的冷冻结缔组织BG5培养基进行实验,观察到的光密度为7.8。优化培养基的成本效益以每克细胞在250毫升Erlenmeyer烧瓶中生产6.25单位价格确定。
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