Abstract
In the present study, conditions for Bacillus mucilaginous fermentation using Agaricus bisporus wastewater as culture medium were optimized. We analyzed the total number of living B. mucilaginous in the fermentation broth using multispectral imaging flow cytometry. Single-factor experiments were carried out, where a Plackett–Burman design was used to screen out three factors from the original six factors of processing wastewater solubility, initial pH, inoculum size, liquid volume, culture temperature, and rotation speed that affected the total number of viable B. mucilaginous. The Box–Behnken response surface method was used to optimize interactions between the three main factors and predict optimal fermentation conditions. Factors significantly affecting the total number of viable B. mucilaginous, including shaking speed, culturing temperature, and initial pH, were investigated. The optimum conditions for B. mucilaginous fermentation in A. bisporus wastewater were a rotational speed of 195 rpm, culture temperature of 29 °C, initial pH of 6.5, solubility of 0.5%, 8% inoculation volume, and 90 mL liquid volume in a 250 mL flask, culture time of 48 h. Under these conditions, the concentration of total viable bacteria reached 2.16 ± 0.02 × 108 Obj/mL, which meets the national standard. A. bisporus wastewater can be used for the cultivation of B. mucilaginous.
Highlights
Agaricus bisporus (Lange) Sing is the most widely cultivated and most consumed edible mushroom with the highest yield in the world, accounting for about 25% of the total world edible fungus production
Detection of the total number of viable bacteria during fermentation A scatter plot comparing fluorescence intensities of SYTO 9 in the green channel and propidium iodide (PI) in the red channel was generated for the in-focus population
In the present study, Plackett–Burman was combined with Box–Behnken experimental design to optimize the culture conditions of B. mucilaginous
Summary
Agaricus bisporus (Lange) Sing is the most widely cultivated and most consumed edible mushroom with the highest yield in the world, accounting for about 25% of the total world edible fungus production. Due to the short storage period of fresh A. bisporus, the main form of international trade is mainly canned processed products. Huang et al AMB Expr (2018) 8:141 as free protein, polysaccharides, mannitol, and mineral ions (Lin et al 2016), so it can be used as a natural medium for the cultivation of certain beneficial bacteria or plants (Zhan et al 2017). If the industrial wastewater produced in the industry of A. bisporus could be used as a natural medium for B. mucilaginous that is one of the most important functional bacteria in microbial fertilizers widely used in the agricultural industry, which would provide theoretical support for microbial fertilizers’ fermentation and the development of the downstream industry of A. bisporus Since B. mucilaginous can transfer the insoluble phosphorus that is present as either an inorganic mineral such as apatite or as one of several organic forms including inositol phosphate (soil phytate), phosphomonesters, and phosphotriesters in the soil to support plant growth, fix nitrogen and provide it to plants (Rojas et al 2001; Javadi Nobandegani et al 2015; Kuan et al 2016), and it generates organic acids, amino acids, polysaccharides, hormones, and other substances absorbed and utilized by plants during growth and reproduction (Glick 2012; Koroney et al 2016; Khalid et al 2017; Schütz et al 2017).
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