Microbial synergy between Pichia kudriazevii YS201 and Bacillus subtilis BS38 improves pulp degradation and aroma production in cocoa pulp simulation medium

摘要

Interactions between two major microorganisms from Ivorian cocoa fermentation, namely Bacillus subtilis BS38 and Pichia kudriazevii YS201, were investigated during fermentation in cocoa pulp simulation medium. The strains were mutually inhibitory, with Bacillus being more susceptible to this antagonistic effect than Pichia. However, both strains yielded different pulp-degrading enzymes, namely polygalacturonase (PG) from Pichia and pectate lyase (Pel) from Bacillus, that cooperate to efficiently breakdown pectin and vegetable pulp. The quantification of aromas from microbial cultures using Gas Chromatography-Mass Spectroscopy (GC-MS) coupled with headspace microextration (SPME) method, showed that P. kudriazevii produce mainly alcohols such as ethanol (63.165 g/L), phenylethanol (1.005 g/L), methylbutanol (0.138 g/L) and esters, notably ethyl acetate (0.037 g/L) and isoamyl acetate (0.032 g/L). The volatile fraction produced by Bacillus was dominated by butanediol (5.707 g/L), acetoin (1.933 g/L), phenylethanol (0.035 g/L) and acetic acid (0.034 g/L). In co-culture, Bacillus produced low levels of aroma compounds whereas a moderate decrease in the production of these compounds was observed in the yeasts strain. Thus, the dominant aromas present in the co-culture were mainly those from the yeasts strain; however, a 1.37 fold increase of ethanol production was observed in co-culture indicating a synergy between the strains. This study showed that cooperation between B. subtilis BS38 and P. kudriazevii YS201 leads principally to increasing pulp degradation and ethanol production, known as desirable properties for a well processing of cocoa fermentation.