Optimization of rice wine fermentation process based on the simultaneous saccharification and fermentation kinetic model

Chinese rice wine making is a typical simultaneous saccharification and fermentation(SSF) process.During the fermentation process,temperature is one of the key parameters which decide the quality of Chinese rice wine.To optimize the SSF process for Chinese rice wine brewing,the effects of temperature on the kinetic parameters of yeast growth and ethanol production at various temperatures were determined in batch cultures using a mathematical model.The kinetic parameters as a function of temperature were evaluated using the software Origin8.0.Combing these functions with the mathematical model,an appropriate form of the model equations for the SSF considering the effects of temperature were developed.The kinetic parameters were found to fit the experimental data satisfactorily with the developed temperature-dependent model.The temperature profile for maximizing the ethanol production for rice wine fermentation was determined by genetic algorithm.The optimum temperature profile began at a low temperature of 26 °C up to 30 h.The operating temperature increased rapidly to 31.9 °C,and then decreased slowly to 18 °C at 65 h.Thereafter,the temperature was maintained at18 °C until the end of fermentation.A maximum ethanol production of 89.3 g·L^(-1)was attained.Conceivably,our model would facilitate the improvement of Chinese rice wine production at the industrial scale.

Identification of acetic-acid tolerance of Saccharomyces cerevisiae strains by microsatellite markers

The aim of this paper is to use the microsatellites to evaluate acid-tolerance in Saccharomyces(S.) cerevisiae. Microsatellites have been widely used as the molecular marker to classify and identify S. cerevisiae strains, analyze genetic relationships among strains, and reveal genetic diversity of S. cerevisiae populations. In this paper, 25 key microsatellites of S. cerevisiae from 44 industrial yeast strains are investigated in the medium withconcentration gradients of acetic acid. Based on the analysis of correlations between the key microsatellite loci repeat numbers and acid-tolerance of the strains, the allele size of 4P14 a and 10P13 is positively related to acid-tolerance(p ? 0.05), the allele size of AT-X, 4P1 a and 10P8 are significantly negatively related to acid-tolerance(p ? 0.01). The above results provide informations on the molecular biodiversity of S. cerevisiae strains and can be a theoretical guidance for molecular marker assisted selection.

Highly efficient synthesis of(R)-1,3-butanediol via anti-Prelog reduction of 4-hydroxy-2-butanone with absolute stereoselectivity by a newly isolated Pichia kudriavzevii

(R)-1,3-butanediol is an important pharmaceutical intermediate, and the synthesis of(R)-1,3-butanediol using green biological methods has recently been of interest for industrial application. Here, a novel strain QC-1 that efficiently transforms 4-hydroxy-2-butanone to(R)-1,3-butanediol was isolated from soil samples. Based on morphological, physiological, and biochemical tests and 5.8 S-internal transcribed spacer sequencing, the strain was identified as Pichia kudriavzevii QC-1. The reaction conditions were optimized to 35 ℃, pH 8.0, rotation speed 200 rpm, and 6:5 mass ratio of glucose to 4-hydroxy-2-butanone. Evaluation of the effects of 4-hydroxy-2-butanone concentrations on yield and cell survival rate showed that 85.60 g·L^-1 product accumulated, with an enantiomeric excess of more than 99%, when 30 g·L^-14-hydroxy-2-butanone was added at 0, 10, and 30 h in a 3-L bioreactor. Thus, strain QC-1 showed excellent catalytic activity and stereoselectivity for the synthesis of(R)-1,3-butanediol from 4-hydroxy-2-butanone.

Response of peroxidase and catalase to acid rain stress during seed germination of rice,wheat,and rape

Seed germination of plants with various acid-resis tance display different responses to acid rain.To understand the reason why such differences occur,the effects of sim ulated acid rain(pH 2.5-5.0)on the activities of peroxidase(POD)and catalase(CAT)during sced ger-mination of rice(O.sativa),wheat(T.aestivurm),and rape(B.chinensis var.oleifera)were investigated.R cesults indi-cated that the maximum change in activities of CAT and POD by acid rain treatment with different acidity and time in relation to the referent treatment without acid rain,was in the order:rice(28.8%,31.7%)<wheat(34.7%,48.3%)<rape(79.3%,50.0%).The pH level for which the treatment with acid rain did not cause signif-cant difference(p<0.05)was in the order:rice(3.5)>wheat(4.0)>rape(5.0).Moreover,the change in activity of POD was higher than that of CAT,which showed that POD was more sensitive to acid rain stress than CAT.The difference in the ability of POD and CAT in removing free radicals was one reason why the germina-tion indexes of these three species behaved differently.

Distinct co‑occurrence patterns and driving forces of abundant and rare bacterial communities in the multispecies solid‑state fermentation process of cereal vinegar

Multispecies solid-state fermentation is a traditional processing technique for the traditional Chinese food,such as cereal vinegar,Baijiu,etc.Generally,few abundant and many rare microbes were involved in such processes,and the necessity and roles of the latter are less studied.Here the co-occurrence patterns of abundant and rare bacterial community and abiotic factors infuencing their community assembly were investigated in acetic acid fermentation following starter inoculation,using Zhenjiang aromatic vinegar as a model system.Abundant taxa that contribute to the function of accumulating acid exhibited a ubiquitous distribution while the distribution of rare taxa along the fermentation process unraveled.The species composition of the rare taxa signifcantly altered,but abundant taxa were maintained after inoculation.Moreover,the diversity of rare taxa changed more signifcantly than that of abundant taxa.Both abundant and rare sub-communities,which were contributed more with species turnover than species richness,were demonstrated to be driven by pH,acetic acid,ammonium nitrogen,and ethanol.Stochastic processes regulated the assembly of both sub-communities,but more prominent on rare sub-communities.Co-occurrence network was more governed by rare sub-communities,and the co-variations between microbial communities were predominantly positive,implying that rare taxa played more important role in the fermentation stability and network robustness.Furthermore,seven network connectors were identifed,and three of them belonged to rare taxa.These microbes of diferent modules were enriched at particular phases of fermentation.These results demonstrate the ecological signifcance of rare bacteria and provide new insights into understanding the abiotic factors infuence microbial structure in traditional fermented foods.

Enzymatic cascade systems for D-amino acid synthesis:progress and perspectives

D-amino acids,different from the ubiquitous L-amino acids,are recognized as the“unnatural”amino acids.The applications of D-amino acids have drawn increasing interest from researchers in recent years,and D-amino acids are widely used in various industries,including for food products,pharmaceuticals,and agricultural chemicals.Inspired by the prevalent appli-cations,many synthetic methods for D-amino acids have been developed,which are mainly divided into chemical synthetic methods and biosynthetic methods.Chemical synthesis of D-amino acids has a variety of disadvantages such as multiple reaction steps,low yields,low reaction rates,and difficulties in product extraction.Thus,biosynthetic methods utilizing enzymes are attracting increasing attention because they are more energy-saving and environmentally friendly compared to traditional chemical synthesis.Among all enzymatic methods,multi-enzymatic cascade catalytic methods have significant advantages,such as lower costs,no need for intermediate separation,and higher catalytic efficiency,which is ascribed to the spatial proximity of biocatalysts.In this review,advances in multi-enzyme cascade catalytic systems as well as chemo-enzymatic approaches to synthesize D-amino acids are discussed.

Advances in Fe(II)/2-ketoglutarate-dependent dioxygenase-mediated C-H bond oxidation for regioselective and stereoselective hydroxyl amino acid synthesis:from structural insights into practical applications

The asymmetric hydroxylation of inactive carbon atoms in organic compounds remains an important reaction in the industrial synthesis of valuable chiral compounds.Fe(II)and 2-ketoglutarate-dependent dioxygenases(Fe/2-kg DOs)are the largest known subgroups of mononuclear nonheme-Fe(II)-dependent oxygenases,catalyzing various oxidation reactions of C-H bonds.Recent developments in Fe/2-kg DO-related researches have coupled concepts from bioinformatics,synthetic biology,and computational biology to establish effective biotransformation systems.The most well-studied and characterized activ-ity of the Fe/2-kg DOs is substrate hydroxylation,with regard to which mechanistic studies involving the Fe center assist in engineering the protein frameworks of these enzymes to obtain the desired catalytic enhancements.Amino acids are typical substrates of Fe/2-kg DOs and are usually converted into hydroxyl amino acids,which are widely used as intermediates in pharmaceutical and fine chemical industries.Herein,we have reviewed prior structural and mechanistic studies on Fe/2-kg DOs,as well as studies on the Fe/2-kg DO-mediated selective C-H oxidation process for selective hydroxyl amino acid synthesis,which will further our journey along the promising path of building complexity via C-H bond oxidation.Further,new bioinformatics techniques should be adopted with structure-based protein rational design to mine sequence databases and shrink mutant libraries to produce a diverse panel of functional Fe/2-kg DOs capable of catalyzing targeted reactions.