Clostridium luticellarii sp. nov., isolated from a mud cellar used for producing strong aromatic liquors

Abstract

A strictly anaerobic, Gram-stain-positive bacterium, designated FW431 T , was isolated from a mud cellar used for producing strong aromatic Chinese liquors. The strain was able to produce butanoic acid, an important component of the aroma style of Chinese liquors. Cells of strain FW431 T were straight or slightly curved rods with a polar endospore and peritrichous flagella. The major cellular fatty acids (.10 % of the total) were C 16 : 0 , C 18 : 1 v9c and C 18 : 0 . Biolog assays indicated that the strain preferably metabolizes palatinose, L-fucose, b-hydroxybutyric acid, L-rhamnose and a-ketobutyric acid among 95 carbon sources tested. FW431 T was related most closely to Clostridium ljungdahlii DSM 13528 T and Clostridium kluyveri DSM 555 T based on 16S rRNA gene sequence similarities of 95.0 and 94.2 %, respectively. The DNA G+C content of the genomic DNA was 44.4 mol%. Based on the evidence presented here, FW431 T (5CGMCC 1.5201 T 5KCTC 15519 T ) is proposed as the type strain of a novel species, Clostridium luticellarii sp. nov.The strong aroma style Chinese liquors are batch-brewed in mud cellars. During the month-long spontaneous solidstate fermentation involved, such cellars are in an anaerobic condition and contain high ethanol concentrations. The brewing process is able to accumulate diverse anaerobic and facultatively aerobic microbes (Wang et al., 2014). These adaptive micro-organisms are able to produce many low-molecular-mass organic acids, which are esterified to various esters or other volatile compounds, these determining the quality and aromatic style of the liquor (Fan & Xu, 2000;Jia, 2003;Hu et al., 2004;Xu et al., 2010;Zheng et al., 2011). Here we report the isolation and characterization of a strictly anaerobic, Gram-stain-positive bacterium, designated FW431 T , isolated from a mud cellar. 2010) agar plates. The RCM had the following composition (per litre): 5 g glucose, 10 g tryptone, 3 g yeast extract, 5 g beef extract, 1 g soluble starch, 5 g NaCl, 3 g sodium acetate, 5 g L-cystine, 1 mg resazurin and 1.5 % agar. The pH was adjusted to 6.5 before autoclaving. The plates were incubated at 37 8C in an anaerobic system (MART Microbiology) filled with a gas mixture of N 2 /H 2 /CO 2 (8 : 1 : 1). The strain was picked as a single colony and further purified by transferring to fresh RCM agar plates several times. The purified strain was stored under liquid nitrogen in tubes containing 20 % glycerol solution.The 16S rRNA gene sequence was amplified from FW431 T using the method described by Wang et al. (2014). The sequence determined was 1441 bp long. A BLAST search of the 16S rRNA gene sequence against the GenBank database showed that the strain shared highest similarity with Clostridium ljungdahlii DSM 13528 T and Clostridium kluyveri DSM 555 T (95.0 and 94.2 %, respectively). Accordingly, further taxonomic identification was performed with these two reference strains, purchased from the Deutsche Sammlung von Micro-organismen und Zellkulturen GmbH (DSMZ, Braunschweig, German). The Clostridi...

Hydrogen as a Co-electron Donor for Chain Elongation With Complex Communities

“…Moreover, a caproate producer recently isolated in our laboratory, Clostridiales bacterium isolate BL-3, is also closely related to C. luticellarii (Liu et al, 2020b,c). C. luticellarii is closely related to C. ljungdahlii (a solventogen and syngas fermenter) and C. kluyveri (a chain elongator) (Wang et al, 2015;de Smit et al, 2019). Moreover, it is important to highlight that the fact that no ethanol accumulated in our experiments should not exclude the possibility of this ED being the one intermediating the H 2 -to-caproate through interspecies transfer.…”

“…With relatively high abundances in all communities, the genus Clostridium sensu stricto 12 thrived particularly in communities A and C after lactate and ethanol addition (Figure 8). So far, hydrogenotrophy and caproate production have not been reported as functions of C. luticellarii (Wang et al, 2015). However, its genome harbors typical genes of the Wood-Ljungdahl pathway (Poehlein et al, 2018) and C. luticellarii was the main candidate to elongate propionate to valerate and to produce i-butyrate in a recent study (de Smit et al, 2019).…”

Continuous n-valerate formation from propionate and methanol in an anaerobic chain elongation open-culture bioreactor

“…Clostridium luticellarii is a known butyrate-producing strain (Additional file 1: Table S11) within the Clostridium sensu stricto 12 genus. It shares the highest similarities with Clostridium ljungdahlii and Clostridium kluyveri [44]. C. luticellarii is the prime candidate for performing the methanol-based chain elongation, which well fits its similarities to a Wood–Ljungdahl-harbouring ( C. ljungdahlii ) microorganism and a reverse β-oxidation-harbouring ( C. kluyverii ) microorganism.…”

Diversity and Function of Microbial Community in Chinese Strong-Flavor Baijiu Ecosystem: A Review

“…Other dominant genera included Weissella, Pediococcus ( Yang J.-G. et al, 2017 ), Lactobacillus, Acetobacter ( Ming et al, 2013 ) from daqu; and Sporolactobacillus, Clostridium, Mycobacterium , and Flavobacterium from pit mud ( Yue et al, 2007 ). In addition, six new species of bacteria [ Paenibacillus vini ( Chen et al, 2015 ), Bacillus vini ( Ma et al, 2016 ), Lysobacter zhanggongensis ( Zhang et al, 2017 ), Clostridium swellfunianum ( Liu et al, 2014 ), C. luticellarii ( Wang et al, 2015 ), and C. liquoris ( Yin et al, 2016 )] were detected and identified from the SFB ecosystem.…”