Zijun Xiao

Strategies for enhancing fermentative production of acetoin: A review

Acetoin is a volatile compound widely used in foods, cigarettes, cosmetics, detergents, chemical synthesis, plant growth promoters and biological pest controls. It works largely as flavour and fragrance. Since some bacteria were found to be capable of vigorous acetoin biosynthesis from versatile renewable biomass, acetoin, like its reduced form 2,3-butanediol, was also classified as a promising bio-based platform chemical. In spite of several reviews on the biological production of 2,3-butanediol, little has concentrated on acetoin. The two analogous compounds are present in the same acetoin (or 2,3-butanediol) pathway, but their production processes including optimal strains, substrates, derivatives, process controls and product recovery methods are quite different. In this review, the usages of acetoin are reviewed firstly to demonstrate its importance. The biosynthesis pathway and molecular regulation mechanisms are then outlined to depict the principal network of functioning in typical species. A phylogenetic tree is constructed and the relationship between taxonomy and acetoin producing ability is revealed for the first time, which will serve as a useful guide for the screening of competitive acetoin producers. Genetic engineering, medium optimization, and process control are effective strategies to improve productivity as well. Currently, downstream processing is one of the main barriers in efficient and economical industrial acetoin fermentation. The future prospects of microbial acetoin production are discussed in light of the current progress, challenges, and trends in this field.

Thermophilic fermentation of acetoin and 2, 3-butanediol by a novel Geobacillus strain

BackgroundAcetoin and 2,3-butanediol are two important biorefinery platform chemicals. They are currently fermented below 40°C using mesophilic strains, but the processes often suffer from bacterial contamination.ResultsThis work reports the isolation and identification of a novel aerobic Geobacillus strain XT15 capable of producing both of these chemicals under elevated temperatures, thus reducing the risk of bacterial contamination. The optimum growth temperature was found to be between 45 and 55°C and the medium initial pH to be 8.0. In addition to glucose, galactose, mannitol, arabionose, and xylose were all acceptable substrates, enabling the potential use of cellulosic biomass as the feedstock. XT15 preferred organic nitrogen sources including corn steep liquor powder, a cheap by-product from corn wet-milling. At 55°C, 7.7 g/L of acetoin and 14.5 g/L of 2,3-butanediol could be obtained using corn steep liquor powder as a nitrogen source. Thirteen volatile products from the cultivation broth of XT15 were identified by gas chromatography–mass spectrometry. Acetoin, 2,3-butanediol, and their derivatives including a novel metabolite 2,3-dihydroxy-3-methylheptan-4-one, accounted for a total of about 96% of all the volatile products. In contrast, organic acids and other products were minor by-products. α-Acetolactate decarboxylase and acetoin:2,6-dichlorophenolindophenol oxidoreductase in XT15, the two key enzymes in acetoin metabolic pathway, were found to be both moderately thermophilic with the identical optimum temperature of 45°C.ConclusionsGeobacillus sp. XT15 is the first naturally occurring thermophile excreting acetoin and/or 2,3-butanediol. This work has demonstrated the attractive prospect of developing it as an industrial strain in the thermophilic fermentation of acetoin and 2,3-butanediol with improved anti-contamination performance. The novel metabolites and enzymes identified in XT15 also indicated its strong promise as a precious biological resource. Thermophilic fermentation also offers great prospect for improving its yields and efficiencies. This remains a core aim for future work.

Generation of Acetoin and Its Derivatives in Foods

Acetoin is a common food flavor additive. This volatile compound widely exists in nature. Some microorganisms, higher plants, insects, and higher animals have the ability to synthesize acetoin using different enzymes and pathways under certain circumstances. As a very active molecule, acetoin acts as a precursor of dozens of compounds. Therefore, acetoin and its derivatives are frequently detected in the component analysis of a variety of foods using gas chromatography-mass spectrometry. Because of the increasing importance of these compounds, this paper reviews the origins and natural existence of these substances, physiological roles, the biological synthesis pathways, nonenzymatic spontaneous reactions, and the common determination methods in foods. This work is the first review on dietary natural acetoin.

Acetoin Catabolism and Acetylbutanediol Formation by Bacillus pumilus in a Chemically Defined Medium

Background Most low molecular diols are highly water-soluble, hygroscopic, and reactive with many organic compounds. In the past decades, microbial research to produce diols, e.g. 1,3-propanediol and 2,3-butanediol, were considerably expanded due to their versatile usages especially in polymer synthesis and as possible alternatives to fossil based feedstocks from the bioconversion of renewable natural resources. This study aimed to provide a new way for bacterial production of an acetylated diol, i.e. acetylbutanediol (ABD, 3,4-dihydroxy-3-methylpentan-2-one), by acetoin metabolism. Methodology/Principal Findings When Bacillus pumilus ATCC 14884 was aerobically cultured in a chemically defined medium with acetoin as the sole carbon and energy source, ABD was produced and identified by gas chromatography – chemical ionization mass spectrometry and NMR spectroscopy. Conclusions/Significance Although the key enzyme leading to ABD from acetoin has not been identified yet at this stage, this study proposed a new metabolic pathawy to produce ABD in vivo from using renewable resources – in this case acetoin, which could be reproduced from glucose in this study – making it the first facility in the world to prepare this new bio-based diol product.

Thermophilic production of polyhydroxyalkanoates by a novel Aneurinibacillus strain isolated from Gudao oilfield, China

Polyhydroxyalkanoates (PHAs) are usually biosynthesized using mesophilic strains, but the fermentation processes often suffer from bacterial contamination. This work reports the screening of thermophilic bacteria capable of producing PHAs under elevated temperatures to reduce the contamination risk. Strain XH2 was isolated from an oilfield and identified as Aneurinibacillus sp. by morphology, physiological‐biochemical characterization, and 16S rDNA phylogenetic analysis. This strain can produce PHA granules, which was detected by Nile red staining and transmission electron microscopic imaging. At 55 °C, 111.6 mg l−1 of PHA was produced in a fermentation medium containing glucose, peptone, and yeast extract. If peptone was removed from the medium, the yield of PHA would be enhanced by 2.4 times. The main monomers of the PHA product were identified to be 3‐hydroxybutyrate and 3‐hydroxyvalerate with a molar ratio of 17.2:1 by gas chromatography‐mass spectroscopy (GC‐MS) and nuclear magnetic resonance analyses. Two minor homologues, 3‐hydroxyoctanoate, and 3‐hydroxy‐4‐phenylbutanoate, were tentatively identified by GC‐MS as well. This is the first report of thermophilic PHA bacterial producer from the Firmicutes phylum.

Identification of an integron containing the quinolone resistance gene qnrA1 in Shewanella xiamenensis.

This study investigated multidrug resistance in Shewanella xiamenensis isolated from an estuarine water sample in China during 2014. This strain displayed resistance or decreased susceptibility to ampicillin, aztreonam, cefepime, cefotaxime, chloramphenicol, ciprofloxacin, erythromycin, kanamycin and trimethoprim-sulfamethoxazole. The antimicrobial resistance genes aacA3, blaOXA-199, qnrA1 and sul1 were identified by PCR amplification and by sequencing. Pulsed-field gel electrophoresis and DNA hybridization experiments showed that the quinolone resistance gene qnrA1 was chromosomally located. qnrA1 was located in a complex class 1 integron, downstream from an ISCR1, and bracketed by two copies of qacEΔ1-sul1 genes. This integron is similar to In825 with four gene cassettes aacA3, catB11c, dfrA1z and aadA2az. An IS26-mel-mph2-IS26 structure was also detected in the flanking sequences, conferring resistance to macrolides. This is the first identification of the class 1 integron in S. xiamenensis. This is also the first identification of the qnrA1 gene and IS26-mediated macrolide resistance genes in S. xiamenensis. Presence of a variety of resistance genetic determinants in environmental S. xiamenensis suggests the possibility that this species may serve as a potential vehicle of antimicrobial resistance genes in aquatic environments.

A novel 2,3-xylenol-utilizing Pseudomonas isolate capable of degrading multiple phenolic compounds

This work characterized a novel 2,3-xylenol-utilizing Pseudomonas isolate XQ23. From 16S rRNA phylogenetic analysis, XQ23 was found to be a member of the Pseudomonas putida group. Most of its physiological characteristics also shared similarities to P. putida. Phenols were catabolized by the meta-cleavage pathway. The dependence of the specific growth rate on 2,3-xylenol concentration could be well fitted by the Haldane model, with the maximum occurring at the concentration around 180 mg l(-1). Kinetic parameters indicated that XQ23 was sensitive to 2,3-xylenol and had low affinity. Three patterns, i.e. constant, linear decline, and allometric decline, were proposed to describe the biomass yields of phenols during bacterial degradation and XQ23 under 2,3-xylenol culturing conditions followed the allometric pattern. In a mineral-salts medium supplemented with 180 mg l(-1) of 2,3-xylenol as the sole carbon and energy source, over 40% of 2,3-xylenol was turned into CO(2) to provide energy by complete oxidization.

GC–FID determination of tetramethylpyrazine and acetoin in vinegars and quantifying the dependence of tetramethylpyrazine on acetoin and ammonium

2,3,5,6-Tetramethylpyrazine (TMP) is an important health functional composition in vinegars, but there are controversial viewpoints about its formation mechanism and scarce relevant records on TMP content enhancement in vinegar products. In this study, a simple and accurate solvent extraction coupled with GC-FID method was developed for the simultaneous determination of TMP and acetoin in vinegars and 137 worldwide samples were analyzed. Meanwhile, the ammonium contents of all the vinegar samples were determined using the salicylate method. By nonlinear surface fitting, the concentrations of TMP in vinegars were found to follow second-order polynomial models of acetoin and ammonium concentrations. By isotopic tracing using 13C labelled acetoin and 15N ammonium, acetoin and ammonium were deduced to be the precursors of TMP in vinegars.

Complete genome sequence of the novel thermophilic polyhydroxyalkanoates producer Aneurinibacillus sp. XH2 isolated from Gudao oilfield in China

Aneurinibacillus sp. XH2 (CGMCC 1.15535) was isolated from Gudao oilfield in China. It is able to use simple carbon resources to accumulate Polyhydroxyalkanoates (PHAs) in a thermophilic fashion. Here, we describe the genomic features of this strain. The total genome size of Aneurinibacillus sp. XH2 is 3,664,835bp and contains 3441 coding sequences and 114 tRNAs. The annotated genome sequence of this strain provides the genetic basis for revealing its role as a themophilic PHAs producing bacterium.

Biodegradation of C 5 -C 8 fatty acids and production of aroma volatiles by Myroides sp. ZB35 isolated from activated sludge

In the effluents of a biologically treated wastewater from a heavy oil-refining plant, C5-C8 fatty acids including pentanoic acid, hexanoic acid, heptanoic acid, octanoic acid, and 2-methylbutanoic acid are often detected. As these residual fatty acids can cause further air and water pollution, a new Myroides isolate ZB35 from activated sludge was explored to degrade these C5-C8 fatty acids in this study. It was found that the biodegradation process involved a lag phase that became prolonged with increasing acyl chain length when the fatty acids were individually fed to this strain. However, when fed as a mixture, the ones with longer acyl chains were found to become more quickly assimilated. The branched 2-methylbutanoic acid was always the last one to be depleted among the five fatty acids under both conditions. Metabolite analysis revealed one possible origin of short chain fatty acids in the biologically treated wastewater. Aroma volatiles including 2-methylbutyl isovalerate, isoamyl 2-methylbutanoate, isoamyl isovalerate, and 2-methylbutyl 2-methylbutanoate were subsequently identified from ZB35 extracts, linking the source of the fruity odor to these esters excreted by Myroides species. To our best knowledge, this is the first finding of these aroma esters in bacteria. From a biotechnological viewpoint, this study has revealed the potential of Myroides species as a promising source of aroma esters attractive for food and fragrance industries.