title: Greater enhancement of Bacillus subtilis spore yields in submerged cultures by optimization of medium composition through statistical experimental designs.
abstract: Bacillus subtilis spore preparations are promising probiotics and biocontrol agents, which can be used in plants, animals, and humans. The aim of this work was to optimize the nutritional conditions using a statistical approach for the production of B. subtilis (WHK-Z12) spores. Our preliminary experiments show that corn starch, corn flour, and wheat bran were the best carbon sources. Using Plackett-Burman design, corn steep liquor, soybean flour, and yeast extract were found to be the best nitrogen source ingredients for enhancing spore production and were studied for further optimization using central composite design. The key medium components in our optimization medium were 16.18 g/l of corn steep liquor, 17.53 g/l of soybean flour, and 8.14 g/l of yeast extract. The improved medium produced spores as high as 1.52 +/- 0.06 x 10(10) spores/ml under flask cultivation conditions, and 1.56 +/- 0.07 x 10(10) spores/ml could be achieved in a 30-l fermenter after 40 h of cultivation. To the best of our knowledge, these results compared favorably to the documented spore yields produced by B. subtilis strains.
authors.affiliation: State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, People's Republic of China.
authors.forename: Zhen-Min
authors.initials: ZM
authors.lastname: Chen
authors.forename: Qing
authors.initials: Q
authors.lastname: Li
authors.forename: Hua-Mei
authors.initials: HM
authors.lastname: Liu
authors.forename: Na
authors.initials: N
authors.lastname: Yu
authors.forename: Tian-Jian
authors.initials: TJ
authors.lastname: Xie
authors.forename: Ming-Yuan
authors.initials: MY
authors.lastname: Yang
authors.forename: Ping
authors.initials: P
authors.lastname: Shen
authors.forename: Xiang-Dong
authors.initials: XD
authors.lastname: Chen
mesh_terms: D001412:Bacillus subtilis / Q000254:growth & development / Q000502:physiology*; D001431:Bacteriological Techniques; D019149:Bioreactors; D001709:Biotechnology; D002244:Carbon / Q000378:metabolism; D003470:Culture Media / Q000737:chemistry*; D004043:Dietary Fiber; D005285:Fermentation; D005433:Flour; D005516:Food Microbiology; D008954:Models, Biological; D015233:Models, Statistical; D009584:Nitrogen / Q000378:metabolism; D012107:Research Design; D013025:Glycine max; D013171:Spores, Bacterial / Q000502:physiology*; D003313:Zea mays
chemical_list: D003470:Culture Media; D004043:Dietary Fiber; D002244:Carbon; D009584:Nitrogen