Pilot-scale production of Bacillus subtilis MSCL 897 spore biomass and antifungal secondary metabolites in a low-cost medium

IF 2 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biotechnology Letters Pub Date : 2024-04-12 DOI:10.1007/s10529-024-03481-4

Emils Bolmanis, Oskars Grigs, Elina Didrihsone, Maris Senkovs, Vizma Nikolajeva

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引用次数: 0

Abstract

Objectives

Bacillus subtilis is a plant growth promoting bacterium (PGPB) that acts as a microbial fertilizer and biocontrol agent, providing benefits such as boosting crop productivity and improving nutrient content. It is able to produce secondary metabolites and endospores simultaneously, enhancing its ability to survive in unfavorable conditions and eliminate competing microorganisms. Optimizing cultivation methods to produce B. subtilis MSCL 897 spores on an industrial scale, requires a suitable medium, typically made from food industry by-products, and optimal temperature and pH levels to achieve high vegetative cell and spore densities with maximum productivity.

Results

This research demonstrates successful pilot-scale (100 L bioreactor) production of a biocontrol agent B. subtilis with good spore yields (1.5 × 10⁹ spores mL⁻¹) and a high degree of sporulation (>80%) using a low-cost cultivation medium. Culture samples showed excellent antifungal activity (1.6–2.3 cm) against several phytopathogenic fungi. An improved methodology for inoculum preparation was investigated to ensure an optimal seed culture state prior to inoculation, promoting process batch-to-batch repeatability. Increasing the molasses concentration in the medium and operating the process in fed-batch mode with additional molasses feed, did not improve the overall spore yield, hence, process operation in batch mode with 10 g molasses L⁻¹ is preferred. Results also showed that the product quality was not significantly impacted for up to 12 months of storage at room temperature.

Conclusion

An economically-feasible process for B. subtilis-based biocontrol agent production was successfully developed at the pilot scale.

Abstract Image

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在低成本培养基中试生产枯草芽孢杆菌 MSCL 897 孢子生物量和抗真菌次生代谢物

目标枯草芽孢杆菌是一种植物生长促进菌（PGPB），可作为微生物肥料和生物防治剂，具有提高作物产量和改善养分含量等功效。它能同时产生次级代谢产物和内生孢子，增强了在不利条件下的生存能力，并能消灭竞争微生物。结果这项研究证明，使用低成本的培养基，成功地以中试规模（100 升生物反应器）生产了一种生物控制剂枯草芽孢杆菌，其孢子产量高（1.5×109 个孢子 mL-1），孢子化程度高（80%）。培养样本对几种植物病原真菌表现出卓越的抗真菌活性（1.6-2.3 厘米）。研究了一种改进的接种物制备方法，以确保接种前种子培养的最佳状态，提高批次间的工艺可重复性。提高培养基中的糖蜜浓度以及在喂料批次模式下使用额外的糖蜜喂料并不能提高总体孢子产量，因此最好在批次模式下使用 10 克糖蜜 L-1 进行工艺操作。结果还显示，在室温下储存 12 个月后，产品质量也没有受到明显影响。

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来源期刊

Biotechnology Letters 工程技术-生物工程与应用微生物

CiteScore

5.90

自引率

3.70%

发文量

108

审稿时长

1.2 months

期刊介绍： Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them. All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included. Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields. The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories. Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.