Fermentation Procedure Research Articles

Study on Quality Traits for Steamed Bread Made from 25 Representative Wheat Varieties in Shandong Province

In order to study steamed bread quality traits and explore good wheat cultivars, 25 wheat cultivars grown in Shandong province were used through one-process fermentation procedure to examine the variability of steamed bread quality traits and the correlation between quality and score. The results showed that all the steamed bread quality traits expressed large variable coefficient except volume, L* value, l* value and cohesiveness traits, which suggested that the genetic diversity made from wheat in Shandong province was large. The average comprehensive score of the steamed bread was 76.8 for the tested 25 varieties, and ten varieties reached the good steamed bread level, which were Liangxing 99, Luyuan 205, Jimai 22, Jinan 17, Tainong 19, Wennong 17, Huaiyin 84137, and Huaimai 8. The main quality factors affecting the steamed bread score were volume, L* value, A* value, hardness, chewiness, and adhesiveness. Among these factors, the adhesiveness has a significantly positive correlation with the hardness and cohesive, and the volume has a significantly positive correlation with the A* value. The volume, L* value has a significantly positive correlation with score, A* value, hardness and chewiness, and gumminess has a significantly negative correlation with the score. There was no correlation between the score and other traits. The results will benefit the understanding of steamed bread quality in wheat cultivated in Shandong. l

Machine Learning of Designed Translational Control Allows Predictive Pathway Optimization in Escherichia coli.

The field of synthetic biology aims to make the design of biological systems predictable, shrinking the huge design space to practical numbers for testing. When designing microbial cell factories, most optimization efforts have focused on enzyme and strain selection/engineering, pathway regulation, and process development. In silico tools for the predictive design of bacterial ribosome binding sites (RBSs) and RBS libraries now allow translational tuning of biochemical pathways; however, methods for predicting optimal RBS combinations in multigene pathways are desirable. Here we present the implementation of machine learning algorithms to model the RBS sequence-phenotype relationship from representative subsets of large combinatorial RBS libraries allowing the accurate prediction of optimal high-producers. Applied to a recombinant monoterpenoid production pathway in Escherichia coli, our approach was able to boost production titers by over 60% when screening under 3% of a library. To facilitate library screening, a multiwell plate fermentation procedure was developed, allowing increased screening throughput with sufficient resolution to discriminate between high and low producers. High producers from one library did not translate during scale-up, but the reduced screening requirements allowed rapid rescreening at the larger scale. This methodology is potentially compatible with any biochemical pathway and provides a powerful tool toward predictive design of bacterial production chassis.

Induced expression of the alcohol acetyltransferase gene ATF1 in industrial yeast Saccharomyces pastorianus TUM 34/70.

Targeted induced gene expression for industrial fermentation processes in food and beverage production could fulfill future demands. To avoid metabolic burden and disturbances owing to the fermentation procedure, induced gene expression is necessary for combating stress, such as that caused by temperature shifts that occur during the transition from fermentation to maturation in the brewing process. The aim of this study was to target gene expression in industrial yeast using stress-responsive promoters and homologues of the selection marker SMR1. Self-cloning strains of the industrial brewing yeast Saccharomyces pastorianus TUM 34/70 were constructed to overexpress the alcohol acetyltransferase (ATF1) gene under the control of inducible promoters P SSA3, P HSP104 and P UBI4. Transcription analysis shows the highest induction after 72h of shock situation for P HSP104 with 1.3-fold and P UBI4 with 2.2-fold. Further, at the end of shock situation the concentrations of ethyl acetate were 1.2- and 1.3-fold higher than the wild type for P HSP104 and P UBI4, respectively. In addition, the influence of the final temperature and temporal sequence of temperature shock to 4°C had a major impact on expression patterns. Therefore, these data show that temperature-induced gene expression of self-cloning industrial yeast could be an option for optimization of the beverage fermentation.

Evaluation of ethyl carbamate formation in Luzhou-flavor spirit during distillation and storage processes

Ethyl carbamate (EC) is a toxic contaminant that poses hazard to humans due to its potential carcinogenicity. This compound has been widely detected in fermented foods and alcoholic beverages such as Chinese spirits. Clarifying the mechanism of EC formation is difficult but necessary for reducing EC levels in Chinese spirits because of their complex fermentation procedures. The formation of ethyl carbamate during distillation and spirit aging were evaluated in this study. All grains used as materials for producing Luzhou-flavor spirit contained ethyl carbamate precursors, urea and citrulline. The highest levels of urea (93 μg/kg) and citrulline (83 μg/kg) were detected in wheat and sorghum, respectively. EC and its precursors were introduced into the raw spirit of Luzhou-flavor spirit during distillation. EC, which was present at 92 μg/l concentration in raw spirit, came from the distillation process. EC content in the raw spirit increased to 66% during the first year of storage because of spontaneous formation of EC from its precursors mainly urea in the raw spirit.

Integrated production of second generation ethanol and lactic acid from steam-exploded elephant grass

Elephant grass was subjected to steam explosion to enhance cellulose accessibility and convert it into ethanol. After catalyzed pretreatment at 190 °C for 5 min, enzymatic hydrolysis was carried out using high rate of solid loading combined with different enzyme dosages. Assays employing 20% (w/v) solids loading and an enzyme dosage of 20 FPU g−1 substrate led to a yield of 86.02 g glucose released per 100 g potential glucose in the water insoluble solids. This condition was selected to carry out the simultaneous saccharification and fermentation procedure through S. cerevisiae CAT-1, producing 42.25 g L−1 ethanol with a yield of 74.57% regard to the maximum theoretical. The liquor containing C5 and C6-sugars was successfully converted into lactic acid using L. buchneri NRRL B-30929, resulting in 13.35 g L−1 with a yield of 68.21% in relation to the maximum theoretical.

Microbial Diversity of Commercial Makgeolli and Its Influence on the Organoleptic Characteristics of Korean Rice Sourdough, Jeung-Pyun.

Sourdough is made by fermentation of dough by lactic acid bacteria (LAB) and yeast to improve bread properties like volume, flavor, and texture. A Korean traditional sourdough was made by fermenting rice flour with rice wine (makgeolli) and used to make sponge-like bread (jeung-pyun). The aim of this study was to investigate the microbial diversity of makgeolli products and their influence on the organoleptic quality of jeung-pyun. Three commercial makgeolli were tested for jeung-pyun production, with each product exhibiting varied dough swelling rates and organoleptic qualities, and among them, J-product was ranked highest in texture and taste. Microbial analysis of the three makgeolli also showed a big difference in their population and diversity. J-product had the highest LAB and yeast counts, and the predominant species were Lactobacillus casei, Lactobacillus brevis, Leuconostoc pseudomenteroides, and Saccharomyces cerevisiae. Using J-product, sourdough was fermented at 25°C, 30°C, and 35°C, and the microbial growth in and textural properties of jeung-pyun were examined by instrumental and sensory tests. At high temperature (35°C), the rates of dough swelling and acidification were fast due to rapid microbial growth mainly caused by LAB, resulting in a short leavening time and soft and sour jeung-pyun. Sensory tests showed consumer preference for the soft and mild-sour jeung-pyun. This study shows that LAB in makgeolli play key roles in production of jeung-pyun, influencing the textural and sensory properties. For the production of high-quality jeung-pyun, development of LAB starters with high gas productivity and low acidity and establishment of an optimal fermentation procedure for rice dough are necessary.

Global substitution of hemeproteins with noncanonical amino acids in Escherichia coli with intact cofactor maturation machinery.

Global substitution of canonical amino acids (cAAs) with noncanonical (ncAAs) counterparts in proteins whose function is dependent on post-translational events such as cofactor binding is still a methodically challenging and difficult task as ncAA insertion generally interferes with the cofactor biosynthesis machinery. Here, we report a technology for the expression of fully substituted and functionally active cofactor-containing hemeproteins. The maturation process which yields an intact cofactor is timely separated from cAA→ncAA substitutions. This is achieved by an optimised expression and fermentation procedure which includes pre-induction of the heme cofactor biosynthesis followed by an incorporation experiment at multiple positions in the protein sequence. This simple strategy can be potentially applied for engineering of other cofactor-containing enzymes.

Validation of Fermentation and Processing Procedures for the Commercial-Scale Production of a Live, Attenuated Edwardsiella ictaluri Vaccine for Use in Channel Catfish Aquaculture.

Mortality associated with Edwardsiella ictaluri infection is a serious impediment to the commercial production of fingerling Channel Catfish Ictalurus punctatus. A patented, live, attenuated, orally delivered vaccine has been developed that offers exceptional protection against E. ictaluri infection in both laboratory and small-scale pond trials. Further vaccine development is contingent on the successful completion of large-scale field trials that accurately reflect industry conditions. This current work focuses on the validation of fermentation protocols and the optimization of downstream processing procedures to produce sufficient quantities of vaccine to conduct commercial-scale field trials. Eight vaccine serials were produced from a master seed stock (S97-773-340X2) in a 50-L floor model fermenter over two consecutive years. Following fermentation, cells were harvested, concentrated 10-fold, and cryogenically stored (-74°C). To assess processing protocols and determine shelf life of cryogenically stored vaccine, serials were tested for cell viability and vaccine potency at various intervals over 24 months. There were no significant differences in cell viability between the fresh vaccine and the stored frozen product. All serials provided a high level of protection (77-100% relative percent survival) against E. ictaluri infection in juvenile Channel Catfish and exhibited excellent poststorage viability. This data demonstrates that the live, attenuated, orally delivered vaccine can be stored at -74°C for at least 2 years with no reduction in cell viability or vaccine potency. Received May 17, 2016; accepted January 19, 2017.

Effects of Fermented Edible Seeds and Their Products on Human Health: Bioactive Components and Bioactivities.

There is a long history of using fermentation in food production. Edible seeds, such as certain beans and cereal grains, are important in the human diet and provide many health benefits. Various microbes, such as lactic acid bacteria, molds, and yeasts, considered as generally recognized as safe (GRAS) microbes, are commonly used to ferment edible seeds and their products. Fermentation can change bioactive components and produce new bioactivities. In order to highlight the importance of fermentation on bioactive components and bioactivities in edible seeds, this review, therefore, summarizes recent relevant studies and discusses fermentation procedures and influences of fermentation on their bioactive components and bioactivities. Overall, fermented edible seeds and their products contain enhanced bioactive components, especially γ-aminobutyric acid and natural phenolics, and they possess versatile bioactivities, such as antioxidant and anti-cancer effects, and, therefore, can be recommended as an important part of the human diet, or they can be developed into functional foods to help in the prevention of certain chronic diseases.

Model-based characterisation of growth performance and l-lactic acid production with high optical purity by thermophilic Bacillus coagulans in a lignin-supplemented mixed substrate medium

Three Bacillus coagulans strains were characterised in terms of their ability to grow in lignin-containing fermentation media and to consume the lignocellulose-related sugars glucose, xylose, and arabinose. An optical-density high-throughput screening was used for precharacterisation by means of different mathematical models for comparison (Logistic, Gompertz, Baranyi, Richards & Stannard, and Schnute). The growth response was characterised by the maximum growth rate and lag time. For a comparison of the screening and fermentation results, an unstructured mathematical model was proposed to characterise the lactate production, bacterial growth and substrate consumption. The growth model was then applied to fermentation procedures using wheat straw hydrolysates. The results indicated that the unstructured growth model can be used to evaluate lactate producing fermentation. Under the experimental fermentation conditions, one strain showed the ability to tolerate a high lignin concentration (2.5g/L) but lacked the capacity for sufficient pentose uptake. The lactate yield of the strains that were able to consume all sugar fractions of glucose, xylose and arabinose was ∼83.4%. A photometric measurement at 280nm revealed a dynamic change in alkali-lignin concentrations during lactate producing fermentation. A test of decolourisation of vanillin, ferulic acid, and alkali-lignin samples also showed the decolourisation performance of the B. coagulans strains under study.

Preparation, structural characterisation and release study of novel hybrid microspheres entrapping nanoselenium, produced by green synthesis.

The main goal of this study was to synthesise and characterise different formulations based on alginate and alginate/chitosan microspheres containing nanoselenium (nano-Se) for controlled delivery applications. Nanosize elemental selenium was produced by using probiotic yogurt bacteria (Lactobacillus casei) in a fermentation procedure. The structural and morphological characterisation of the microspheres was performed by Fourier transform infrared (FTIR), X-ray diffraction (XRD) and scanning electron microscopy (SEM) analysis. FTIR and XRD pattern indicated that was an effective cross-linking of selenium nanoparticles within the polymeric matrix in both cases. The SEM images reveal that selenium nanoparticles are mainly exposed on the surface of alginate, in contrast to porous structure of alginate/chitosan/nano-Se, interconnected in a regular network. This architecture type has a considerable importance in the delivery process, as demonstrated by differential pulse voltammetry. Selenium release from both matrices is pH sensitive. Moreover, chitosan blended with alginate minimise the release of encapsulated selenium, in simulated gastric fluid, and prolong the duration of release in intestinal fluid. The overall effect is the enhancement of total percentage release concomitant with the longer duration of action. The authors' formulation based on alginate/chitosan is a convenient matrix to be used for selenium delivery in duodenum, caecum and colon.

Fermentation strategy to produce high gluconate vinegar

Gluconic acid is a non-volatile acid that has many applications in food, pharmaceutical and cleaning fields. Gluconic acid has been detected as main oxidation product of <em>Acetobacter</em> and <em>Gluconobacter</em> strains growing on grape must, and it plays an important role in Traditional Balsamic Vinegar. Commonly, high gluconate vinegars have a greater physical stability and a greater preference by consumers because are perceived less pungent. In fact, gluconic acid reduces the pH and increases fixed acidity of the vinegar without increasing the sensation of pungency typical of acetic acid. Its taste is acid but mild sweet and, therefore, gluconic acid has influence on the sensory complexity of the vinegar. The aim of this work is to set up a fermentation procedure that improves the quality of balsamic vinegar by using selected yeasts and acetic acid bacteria strains able to oxidize glucose in grape must-based media having a different sugars concentration. In particular, <em>Saccharomycodes</em> <em>ludwigii</em> UMCC 297 and <em>Acetobacter</em> <em>pasteurianus</em> UMCC 1754 strains were chosen as selected starter cultures for small-scale fermentation of cooked grape must, to evaluate the physical-chemical parameters affecting gluconic acid production in the obtained vinegar. The strains used and the control of all production process have been fundamental for obtaining the vinegar with the desired characteristics.

Varieties, production, composition and health benefits of vinegars: A review.

Vinegars are liquid products produced from the alcoholic and subsequent acetous fermentation of carbohydrate sources. They have been used as remedies in many cultures and have been reported to provide beneficial health effects when consumed regularly. Such benefits are due to various types of polyphenols, micronutrients and other bioactive compounds found in vinegars that contribute to their pharmacological effects, among them, antimicrobial, antidiabetic, antioxidative, antiobesity and antihypertensive effects. There are many types of vinegars worldwide, including black vinegar, rice vinegar, balsamic vinegar and white wine vinegar. All these vinegars are produced using different raw materials, yeast strains and fermentation procedures, thus giving them their own unique tastes and flavours. The main volatile compound in vinegar is acetic acid, which gives vinegar its strong, sour aroma and flavour. Other volatile compounds present in vinegars are mainly alcohols, acids, esters, aldehydes and ketones. The diversity of vinegars allows extensive applications in food.

High-throughput fermentation screening for the yeast Yarrowia lipolytica with real-time monitoring of biomass and lipid production.

BackgroundBecause the model yeast Yarrowia lipolytica can synthesize and store lipids in quantities up to 20 % of its dry weight, it is a promising microorganism for oil production at an industrial scale. Typically, optimization of the lipid production process is performed in the laboratory and later scaled up for industrial production. However, the scale-up process can be complicated by genetic modifications that are optimized for one set of growing conditions can confer a less-than-optimal phenotype in a different environment. To address this issue, small cultivation systems have been developed that mimic the conditions in benchtop bioreactors. In this work, we used one such microbioreactor system, the BioLector, to develop high-throughput fermentation procedures that optimize growth and lipid accumulation in Y. lipolytica. Using this system, we were able to monitor lipid and biomass production in real time throughout the culture duration.ResultsThe BioLector can monitor the growth of Y. lipolytica in real time by evaluating scattered light; this produced accurate measurements until cultures reached an equivalent of OD600nm = 115 and a cell dry weight of 100 g L−1. In addition, a lipid-specific fluorescent probe was applied which reliably monitored lipid production up to a concentration of 12 g L−1. Through screening various growing conditions, we determined that a carbon/nitrogen ratio of 35 was the most efficient for lipid production. Further screening showed that ammonium chloride and glycerol were the most valuable nitrogen and carbon sources, respectively, for growth and lipid production. Moreover, a carbon concentration above 1 M appeared to impair growth and lipid accumulation. Finally, we used these optimized conditions to screen engineered strains of Y. lipolytica with high lipid-accumulation capability. The growth and lipid content of the strains cultivated in the BioLector were compared to those grown in benchtop bioreactors.ConclusionTo our knowledge, this is the first time that the BioLector has been used to track lipid production in real time and to monitor the growth of Y. lipolytica. The present study also showed the efficacy of the BioLector in screening growing conditions and engineered strains prior to scale-up. The method described here could be applied to other oleaginous microorganisms.

Effect of fermentation and sterilization on anthocyanins in blueberry.

Blueberry products have various health benefits due to their high content of dietary anthocyanins. The aim of this study was to investigate the impact of fermentation and sterilization on total anthocyanin content, composition and some quality attributes of blueberry puree. The blueberry puree used here was fermented for 40 h at 37 °C by Lactobacillus after sterilization. The method of ultra-performance liquid chromatography-mass spectrometry was optimized for the rapid analysis of anthocyanins. Quality attributes including pH, color, total soluble solids and viscosity were measured. A total of 21 anthocyanins and five anthocyanidins were quantified by ultra-performance liquid chromatography. Fermented blueberry had reduced total anthocyanin content (29%) and levels of individual anthocyanins compared with fresh blueberry. Total anthocyanin content was decreased 46% by sterilization, and different degradation behavior of individual anthocyanin was appeared between fermented and sterilized-fermented blueberry puree. Fermentation and sterilization decreased the total soluble solids and pH and changed color parameters, while minimally influencing viscosity. The loss of total anthocyanin content by fermentation was related to the unstable structure of blueberry anthocyanins. Anthocyanins are sensitive to temperature (>80 °C), and degradation of anthocyanins by sterilization in blueberry should be considered in the fermentation procedure. © 2016 Society of Chemical Industry.

Efficient production of nonactin by Streptomyces griseus subsp. griseus.

Here we report the production of the cyclic macrotetrolide nonactin from the fermentation culture of Streptomyces griseus subsp. griseus. Nonactin is a member of a family of naturally occurring cyclic ionophores known as the macrotetrolide antibiotics. Our fermentation procedure of Streptomyces griseus was performed at 30 °C and 200 rev·min(-1) for 5 days on a rotary shaker. Diaion HP-20 and Amberlite XAD-16 were added to the fermentation medium. Isolated yield of nonactin was up to 80 mg·L(-1) using our methodology. Nonactin is commonly known as an ammonium ionophore and also exhibits antibacterial, antiviral, and antitumor activities. It is also widely used for the preparation of ion-selective electrodes and sensors. Chemical synthesis of nonactin has been achieved by some groups; however, overall yields are very low, making efficient biosynthesis an attractive means of production.

Compositional differences among Chinese soy sauce types studied by 13C NMR spectroscopy coupled with multivariate statistical analysis

Soy sauce a well known seasoning all over the world, especially in Asia, is available in global market in a wide range of types based on its purpose and the processing methods. Its composition varies with respect to the fermentation processes and addition of additives, preservatives and flavor enhancers. A comprehensive 1H NMR based study regarding the metabonomic variations of soy sauce to differentiate among different types of soy sauce available on the global market has been limited due to the complexity of the mixture. In present study, 13C NMR spectroscopy coupled with multivariate statistical data analysis like principle component analysis (PCA), and orthogonal partial least square-discriminant analysis (OPLS-DA) was applied to investigate metabonomic variations among different types of soy sauce, namely super light, super dark, red cooking and mushroom soy sauce. The main additives in soy sauce like glutamate, sucrose and glucose were easily distinguished and quantified using 13C NMR spectroscopy which were otherwise difficult to be assigned and quantified due to serious signal overlaps in 1H NMR spectra. The significantly higher concentration of sucrose in dark, red cooking and mushroom flavored soy sauce can directly be linked to the addition of caramel in soy sauce. Similarly, significantly higher level of glutamate in super light as compared to super dark and mushroom flavored soy sauce may come from the addition of monosodium glutamate.The study highlights the potentiality of 13C NMR based metabonomics coupled with multivariate statistical data analysis in differentiating between the types of soy sauce on the basis of level of additives, raw materials and fermentation procedures.

Biofilm formation on Conservolea natural black olives during single and combined inoculation with a functional Lactobacillus pentosus starter culture

The potential of biofilm formation of multifunctional starters Lactobacillus pentosus B281 and Pichia membranifaciens M3A during inoculated fermentation of Conservolea natural black olives according to Greek-style processing was investigated. Olives were directly brined in 8% (w/v) NaCl following three fermentation procedures namely, i) spontaneous fermentation, ii) inoculated fermentation with L. pentosus B281, and iii) co-inoculated fermentation with L. pentosus B281 and P. membranifaciens M3A. Lactic acid bacteria (LAB) and yeasts were monitored on olives by plate counting for a period of 153 days, whereas the survival of the inoculated strains was confirmed by Pulsed Field Gel Electrophoresis (PFGE) and Restriction Fragment Length Polymorphism (RFLP) analysis. Inoculated fermentation with L. pentosus B281 with/without the presence of the yeast resulted in higher acidification of the brine compared to the spontaneous process where no indigenous LAB could be enumerated. The population of LAB on olives ranged between 5.5 and 6.5 log CFU/g and it was maintained at higher levels compared to yeasts (3.5–4.5 log CFU/g) throughout the process. PFGE analysis revealed that L. pentosus B281 could successfully colonize the surface of black olives presenting high recovery rate (100%) at the end of fermentation in contrast to P. membranifaciens M3A that was successfully recovered (42%) only after 72 days of the process. The obtained results provide interesting perspectives for the production of natural black olives with functional properties.

Ethanol from a biorefinery waste stream: Saccharification of amylase, protease and xylanase treated wheat bran

Biorefining aims to exploit the full value of plant material by sequentially extracting and valorising its components. Many studies focus on the saccharification of virgin biomass sources, but it may be more efficient to pre-extract high-value components before hydrolysis to fermentable sugars. In the current study, a bran residue from de-starched, protein depleted and xylanase treated wheat bran has been subjected to hydrothermal pretreatment, saccharification and fermentation procedures to convert the residue to ethanol. The most effective pretreatment conditions (>190°C, 10min) and saccharification conditions were identified following bench-scale liquid hot water pretreatment. Pre-extraction of enzymatically-hydrolysable starch and xylan reduced the release of furfural production, particularly when lower pretreatment severities were used. Pilot-scale steam explosion of the lignocellulosic residue followed by cellulase treatment and conversion to ethanol at a high substrate concentration (19%) gave an ethanol titre of ≈25g/L or a yield of 93% of the theoretical maximum.

Effect of Maceration Conditions on Chemical Composition and Colour Characteristics of Merlot Wines

Due to their complex composition, Merlot cultivar red grapes were subjected to maceration-fermentation as an important stage for the natural pigments and aroma compounds extraction. The purpose of this paper was the assessment of three pre-fermentation maceration techniques in order to improve wine's characteristics. For the experiment were used Merlot cultivar red grapes. The grapes were removed from their clusters, crushed and than transferred to three maceration techniques: microwave maceration at 650 W for 15 minutes, thermal maceration by heating the grape grains at 70 ⁰C for 20 minutes and classical maceration for 7 days at 20 ⁰C. During the process of maceration and fermentation all the samples were monitored considering the yeast cell count and alcohol content. The wine samples were evaluated in terms of colour, by spectrophotometric method. The evaluation of technological index, the evolution of the yeast cells and the fermentation parameters from the three samples indicate that the most advantageous fermentation procedure was classical maceration-fermentation.