A new method for screening and culture of Clostridium from pit mud under non-anaerobic conditions

Abstract

Strong-flavor Baijiu (SFB) is produced in complex fermentation in pits under ground. Clostridium producing hexanoic acid plays a key role in the flavor formation of SFB. The screening and culture for Clostridium are very difficult because of its strict anaerobic characteristics. In this study, electric field assisted screening (EFAS) was used to screen Clostridium from pit mud, and electric culture (EC) was used to cultivate Clostridium under non-anaerobic conditions. A strain with a high yield of hexanoic acid was screened and named as Clostridium sp. EFAS6. Under non-anaerobic conditions, it grew rapidly only near the cathode end in the EFAS device because of the low oxidation-reduction potential of that electrode. In the experiment of high-density culture in the EC device, the cell concentration reached 10⁶-10⁷. After energy consumption was calculated, the optimal loading voltage was found to be 10 V. In the application, the broth of Clostridium sp. EFAS6 increased the content of ethyl hexanoic in SFB. Under non-anaerobic conditions, the anaerobe was screened by EFAS and cultivated in high density by EC. The EFAS and EC could also be used for the screening and culture of other anaerobes under non-anaerobic conditions.

Keywords: Clostridium; High-density culture; Non-anaerobic conditions; Pit mud; Screening.