Wine Fermentation Research Articles

Monitoring Site-Specific Fermentation Outcomes via Oxidation Reduction Potential and UV-Vis Spectroscopy to Characterize "Hidden" Parameters of Pinot Noir Wine Fermentations.

Real-time process metrics are standard for the majority of fermentation-based industries but have not been widely adopted by the wine industry. In this study, replicate fermentations were conducted with temperature as the main process parameter and assessed via in-line Oxidation Reduction Potential (ORP) probes and at-line profiling of phenolics compounds by UV-Vis spectroscopy. The California and Oregon vineyards used in this study displayed consistent vinification outcomes over five vintages and are representative of sites producing faster- and slower-fermenting musts. The selected sites have been previously characterized by fermentation kinetics, elemental profile, phenolics, and sensory analysis. ORP probes were integrated into individual fermentors to record how ORP changed throughout the fermentation process. The ORP profiles generally followed expected trends with deviations revealing previously undetectable process differences between sites and replicates. Site-specific differences were also observed in phenolic and anthocyanin extraction. Elemental composition was also analyzed for each vineyard, revealing distinctive profiles that correlated with the fermentation kinetics and may influence the redox status of these wines. The rapid ORP responses observed related to winemaking decisions and yeast activity suggest ORP is a useful process parameter that should be tracked in addition to Brix, temperature, and phenolics extraction for monitoring fermentations.

Spectroscopic investigation of exopolysaccharides purified from Arthrospira platensis cultures as potential bioresources

In this work, we employed Raman and Fourier-transform infrared (FTIR) spectroscopies in order to monitor in real time the exopolysaccharides (EPS) components purified from Arthrospira platensis AICB 49 (previously known as Spirulina sp.) species. Preliminary Raman and IR spectra for EPS obtained following a stepwise ethanol extraction are reported as compared to the dextran molecule, a well-known byproduct of wine fermentation. The Raman spectra were recorded with 2 commonly used excitation wavelengths, 532 nm and 633 nm, respectively. The final aim we pursue is empowering microbiological byproducts, that otherwise generate high disposal costs, which hold great promise as competitive and affordable industrial-grade staple. Once we succeed to tackle the accurate composition of such byproducts and their spectral fingerprint, we will be able to estimate their potential as bioresources. As a complementary technique, FTIR spectroscopy was additionally employed for a better elucidation of the molecular composition of the samples. We comprehensively assigned the recorded Raman and IR bands observed for the EPS extracted from A. platensis by assessing a molecular structure investigation and by corroborating the recorded spectral results with recent studies reporting similar saccharide-species.

Impact of high lipid contents on the production of fermentative aromas during white wine fermentation.

In Cognac, the musts are rich in grape solids and fermentations are usually run with turbidities ranging between 500 and 1500 NTU (nephelometric turbidity unit). These conditions, considered favourable for generating the desired organoleptic profiles of the final Eaux-de-vies, are unusual in winemaking, and, consequently, their impact on yeast metabolism is poorly understood. This study aims to better describe and understand the synthesis of fermentative aromas in such lipid-excess conditions, while integrating the effect of two other very important parameters: the initial concentration of assimilable nitrogen and the temperature of fermentation. To reach this objective, a Box-Behnken design was implemented to describe and model the simple effects of these factors as well as their interactions. Although the lipid concentration was very high, impacts on the production of fermentative aromas were observed. Indeed, high lipid levels promoted the synthesis of higher alcohols. Observing this effect was surprising because there is no metabolic connection between the anabolic pathways of production of these alcohols and the lipid pathway. This effect may be partly explained by impairment in the activity of alcohol acetyl transferases in the presence of lipids, which catalyse the conversion of higher alcohols into the corresponding esters. Therefore, in this study, the negative impact of turbidity was very significant on acetate esters related to the production of acetyl-CoA, which was the main molecule disturbed by the strong presence of lipids. Finally, and more surprisingly, lipid intake did not impact the synthesis of ethyl esters, which depended on the concentration of exogenous lipids. KEY POINTS: • Innovative work on the fermentation of white wine musts with very high lipid contents. • Precise fermentation management and monitoring in Cognac-making conditions. • Experimental design to study the impact of lipids, assimilable nitrogen and temperature on fermentative aroma synthesis.

State-of-the-art in analytical methods for metabolic profiling of Saccharomyces cerevisiae

Saccharomyces cerevisiae has many angles, as a model system for eukaryotic biology, as a powerful industrial tool and as a subject widely studied for multiple purposes. The metabolic pathways of this yeast, its primary and secondary metabolites have been analyzed in the last two decades with several technologies. The major technique used nowadays for metabolic profiling is mass spectrometry, very often hyphenated to chromatography separation methods. Gas chromatography-mass spectrometry has been widely applied to S. cerevisiae yeast samples and fermented wine samples, alongside with liquid chromatography-mass spectrometry for untargeted analysis of a wide range of compounds. Capillary electrophoresis-mass spectrometry has been used for the analysis of charged and highly polar molecules, while direct introduction-mass spectrometry is used for fast global screening based on ultra-high resolution mass spectrometry. Another major methodology applied for the yeast characterization is nuclear magnetic resonance, as it is robust, reproducible, non-destructive and produces spectral information that allows an accurate structural elucidation. Finally, some spectroscopic techniques, such as UV–Vis and infrared spectroscopy, have been applied to S. cerevisiae metabolic profiling. These techniques provide less information than mass spectrometry and nuclear magnetic resonance, but still lead to fast global screening of a mixture or of a whole cell, without extraction. This review summarizes the work and research that contributed to S. cerevisiae metabolic profiling during the two last decades.

Origin, Succession, and Control of Biotoxin in Wine.

Wine is a worldwide alcoholic beverage with antioxidant active substances and complex flavors. Moderate drinking of wine has been proven to be beneficial to health. However, wine has some negative components, such as residual pesticides, heavy metals, and biotoxins. Of these, biotoxins from microorganisms were characterized as the most important toxins in wine. Wine fermentation mainly involves alcoholic fermentation, malolactic fermentation, and aging, which endue wine with complex flavors and even produce some undesirable metabolites. These metabolites cause potential safety risks that are not thoroughly understood. This review aimed to investigate the origin, evolution, and control technology of undesirable metabolites (e.g., ochratoxin A, ethyl carbamate, and biogenic amines) in wine. It also highlighted current wine industry practices of minimizing the number of biotoxins in wine.

Yeast Interactions and Molecular Mechanisms in Wine Fermentation: A Comprehensive Review.

Wine can be defined as a complex microbial ecosystem, where different microorganisms interact in the function of different biotic and abiotic factors. During natural fermentation, the effect of unpredictable interactions between microorganisms and environmental factors leads to the establishment of a complex and stable microbiota that will define the kinetics of the process and the final product. Controlled multistarter fermentation represents a microbial approach to achieve the dual purpose of having a less risky process and a distinctive final product. Indeed, the interactions evolved between microbial consortium members strongly modulate the final sensorial properties of the wine. Therefore, in well-managed mixed fermentations, the knowledge of molecular mechanisms on the basis of yeast interactions, in a well-defined ecological niche, becomes fundamental to control the winemaking process, representing a tool to achieve such objectives. In the present work, the recent development on the molecular and metabolic interactions between non-Saccharomyces and Saccharomyces yeasts in wine fermentation was reviewed. A particular focus will be reserved on molecular studies regarding the role of nutrients, the production of the main byproducts and volatile compounds, ethanol reduction, and antagonistic actions for biological control in mixed fermentations.

Towards Accelerated Autolysis? Dynamics of Phenolics, Proteins, Amino Acids and Lipids in Response to Novel Treatments and during Ageing of Sparkling Wine

Premium sparkling wine produced by the traditional method (analogous to the French méthode champenoise) is characterised by the development of aged wine character as a result of a second fermentation in the bottle with lees contact and lengthy ageing. Treatments (microwave, ultrasound, or β-glucanase enzymes) were applied to disrupt the cell wall of Saccharomyces cerevisiae and added to the tirage liquor for the second fermentation of Chardonnay-Pinot Noir base wine cuvée and compared to a control, to assess effects on the release of phenolics, proteins, amino acids, and lipids at 6, 12 and 18 months post-tirage. General responses to wine ageing included a 60% increase in the total phenolic content of older sparkling wines relative to younger wines and an increase in protein concentration from 6 to 12 months bottle age. Microwave and β-glucanase enzyme treatments of yeast during tirage preparation were associated with a 10% increase in total free amino acid concentration and a 10% increase in proline concentration at 18 months bottle age, compared to control and ultrasound treatment. Furthermore, microwave treatment was associated with elevated asparagine content in wine at 18 months bottle age, relative to the control and the other wines. The β-glucanase enzyme and ultrasound treatments were associated with significant accumulation of total lipids, which were driven by 2-fold increases in the phospholipid and monoacylglycerol components in wine at 18 months bottle age and, furthermore, the microwave treatment was associated with elevated triacylglycerol at 18 months bottle age. This study demonstrates that the use of yeast treatments at the tirage stage of sparkling wine production presents an opportunity to manipulate wine composition.

Influence ofSaccharomyces cerevisiaeautochthonousMQ3strain on terpenes during the alcoholic fermentation of Chardonnay dry white wine

Background and Aims Terpenes are major varietal aroma compounds significantly contributing to wine quality and style. Autochthonous microbial starters can promote the formation and release of the terpenes and thus highlight the terroir typicality of wines. This study compared the physicochemical parameters, changes in the terpene concentration, β-D-glucosidase activity and sensory attributes of Chardonnay wines fermented individually by the Saccharomyces cerevisiae autochthonous MQ3 strain isolated from spontaneous fermented grape must and by three commercial S. cerevisiae strains. Methods and Results The evolution of terpenes during alcoholic fermentation was followed with ultrasound-assisted liquid–liquid extraction combined with GC. The physicochemical parameters of wine fermented by the MQ3 strain were significantly different from those of the wines fermented by other yeasts. The pattern of the terpenes detected, including linalool, α-terpineol, farnesol, geraniol, nerol and citronellol, varied during alcoholic fermentation. The MQ3 strain induced greater accumulation of terpenes with increased concentration of α-terpineol, geraniol and (E,E)-farnesol. Besides, the MQ3 strain exhibited an enhanced ability to produce β-D-glucosidase compared to that of the other three strains. With the exception of α-terpineol and (E,E)-farnesol, there was no significant correlation between the concentration of the other terpenoids and β-D-glucosidase activity as shown by Pearson's correlation analysis. Wine produced with the MQ3 strain was characterised by a distinctive complex flavour with enhanced floral and fruity aromas. Conclusions The autochthonous MQ3 strain had a positive effect on the varietal aroma and sensory quality of the wines. Significance of the Study Due to its aroma enhancing ability, the MQ3 strain could be applied to produce quality Chardonnay wine with terroir typicality.

The influence of selected indigenous yeasts on Pinot Noir wine colour properties.

The use of indigenous selected starters in winemaking is gaining interest due to certain advantages for the sensory quality of the wine. The present work shows the results of a laboratory experiment in which the influence of selected indigenous yeasts on the colour characteristics of Pinot Noir was studied with the use of high hydroxycinnamate decarboxylase activity yeasts. Pichia guilliermondii ZIM624 and Wickerhamomyces anomalus S138 yeasts were used in sequential fermentation with two strains of Saccharomyces cerevisiae, the native ZIM2180 strain and commercial Fermol Premier Cru (FPC). In co-inoculation fermentations, non-Saccharomyces yeasts decreased colour intensity (on average by 25.5%). In wines fermented with ZIM624, the concentration of vinylphenolic pyranoanthocyanins increased (average concentration 1.5mg L-1 ). However, vitisin concentration was significantly higher in S138 + FPC fermentation (1.3mg L-1 and an average of 0.9mg L-1 , respectively). Pinot Noir wines fermented with only ZIM2180 and sequential inoculation of ZIM624 + ZIM2180 resulted in significantly higher colour intensity (6.1± 0.0AU and 4.4± 0.0AU, respectively) and lower wine hue parameters compared to other wines. Sensory evaluation also showed that both wines had the highest perceived colour intensity and purple colour suggesting improvement in wine quality parameters. The results confirmed that selected indigenous starters made out of Saccharomyces and non-Saccharomyces yeasts can alter Pinot Noir wine colour parameters and improve wine colour properties. Those yeasts properties should be investigated prior to the development of new commercial starters but also be considered in large scale spontaneous fermentations of low colour intensity red wines like Pinot Noir. © 2021 Society of Chemical Industry.

Use of Yeast Mannoproteins by Oenococcus oeni during Malolactic Fermentation under Different Oenological Conditions.

Oenococcus oeni is the main agent of malolactic fermentation in wine. This fermentation takes place after alcoholic fermentation, in a low nutrient medium where ethanol and other inhibitor compounds are present. In addition, some yeast-derived compounds such as mannoproteins can be stimulatory for O. oeni. The mannoprotein concentration in wine depends on the fermenting yeasts, and non-Saccharomyces in particular can increase it. As a result of the hydrolytic activity of O. oeni, these macromolecules can be degraded, and the released mannose can be taken up and used as an energy source by the bacterium. Here we look at mannoprotein consumption and the expression of four O. oeni genes related to mannose uptake (manA, manB, ptsI, and ptsH) in a wine-like medium supplemented with mannoproteins and in natural wines fermented with different yeasts. We observe a general gene upregulation in response to wine-like conditions and different consumption patterns in the studied media. O. oeni was able to consume mannoproteins in all the wines. This consumption was notably higher in natural wines, especially in T. delbrueckii and S. cerevisiae 3D wines, which presented the highest mannoprotein levels. Regardless of the general upregulation, it seems that mannoprotein degradation is more closely related to the fermenting medium.

Characterization of Free and Bound Monoterpene Alcohols during Riesling Fermentation

The isomeric nature of monoterpenyl glycosides makes unambiguous identification of intact glycosides difficult. As a result, it is challenging to relate the changes in free monoterpenol concentrations to the corresponding glycosides during wine fermentation and storage. In this study, we isolated and identified linalool, nerol, and geraniol monoterpenyl glycosides fromVitis viniferacv. Riesling grapes through fractionation followed by acid or enzyme hydrolysis. Changes in the composition of identified monoterpenyl glycosides and their respective free volatiles were then monitored during alcoholic fermentations of Riesling juice with four different yeast strains across two successive years. The relative concentrations of the volatiles were monitored by solid-phase microextraction gas chromatography mass spectrometry, while ultrahigh-performance liquid chromatography quadrupole time-of-flight mass spectrometry was used for intact glycosides. Glycoside hydrolysis during fermentation could be related to relative concentrations of the corresponding free aglycones. However, other sources of free monoterpenols were also observed. Differences in glycoside hydrolysis among yeast strains and across years were observed and may be related to grape maturity and/or nutrient levels.

Influence of Autochthonous and Commercial Yeast Strains on Fermentation and Quality of Wines Produced from Vranec and Cabernet Sauvignon Grape Varieties from Tikveš Wine-Growing Region, Republic of North Macedonia

Two autochthonous yeast strains called F-8 and F-78 (isolated and selected from the Tikveš wine-producing region) were inoculated in wine musts from Vranec and Cabernet Sauvignon grape varieties. The fermentation process and quality of the produced wines were compared to the wines produced from the same grape varieties, but with a commercial yeast strain (D-80). The fermentation process was undertaken at 23–25 °C for 16 days. The highest alcohol content was detected in Vranec and Cabernet Sauvignon wines fermented with autochthonous F-8 yeast strain. Conversely, Vranec wine fermented by the F-78 yeast strain contained the lowest alcohol content (14.11%). The low sugar amount (2–3 g/L) in both grape varieties is indicated the production of dry wines. The total amount of phenolic compounds was significantly higher in Vranec wines. Unlike the alcohol content, Vranec wine produced by commercial yeast strain D-80 indicated the highest concentration of total phenolic compounds (1450 mg/L) and total anthocyanins (572 mg/L), while the lowest concentrations were observed in wine fermented by autochthonous yeast strain F-78 (1612 mg/L and 470 mg/L, respectively). The maximum color intensity was measured in the Vranec wine fermented by autochthonous yeast strain F-8, while wines from Cabernet Sauvignon grape variety fermented with the same yeast strain F-8 and commercial yeast strain D-80 had comparable values in terms of color intensity.

How to modulate the formation of negative volatile sulfur compounds during wine fermentation?

ABSTRACTBeyond the production of positive aromas during alcoholic fermentation, Saccharomyces cerevisiae metabolism also results in the formation of volatile compounds detrimental to wine quality, including a wide range of volatile sulfur compounds (VSCs). The formation of these VSCs during wine fermentation is strongly variable and depends on biological and environmental factors. First, the comparison of the VSCs profile of 22 S. cerevisiae strains provided a comprehensive overview of the intra-species diversity in VSCs production: according to their genetic background, strains synthetized from 1 to 6 different sulfur molecules, in a 1- to 30-fold concentration range. The impact of fermentation parameters on VSCs production was then investigated. We identified yeast assimilable nitrogen, cysteine, methionine and pantothenic acid contents – but not SO2 content – as the main factors modulating VSCs production. In particular, ethylthioacetate and all the VSCs deriving from methionine catabolism displayed a maximal production at yeast assimilable nitrogen concentrations around 250 mg/L; pantothenic acid had a positive impact on compounds deriving from methionine catabolism through the Ehrlich pathway but a negative one on the production of thioesters. Overall, these results highlight those factors to be taken into account to modulate the formation of negative VSCs and limit their content in wines.

Biogeographical Regionalization of Wine Yeast Communities in Greece and Environmental Drivers of Species Distribution at a Local Scale.

Recent research has expanded our understanding on vineyard-associated fungal community assembly, suggesting non-random distribution and implicating regional differences in the wine terroir effect. Here, we focused on the culturable fraction of the fungal community that resides on grapes and determine wine quality, the so-called wine yeast populations. We aimed to analyze local-scale yeast community assemblages and to test whether the hypothesis of biogeographical patterns also applies to wine yeasts in particular. Surveying 34 vineyards across four main viticultural zones in Greece showed significant trends in vineyard-specific patterns. At a local scale, viticultural regions were also linked to distinct yeast community compositions. Importantly, major yeast populations directly related to wine fermentation contributed significantly to the delimitation of regions, highlighting their potential influence on the regionality of wine characteristics. In terms of the microbial terroir influence, yeast communities within an area were temporarily stable, which is critical for the regional character of the wine. Community structure could be explained only partially by environmental features. Maximum temperature, elevation, and net precipitation were the highest correlated variables with the yeast community biogeographic patterns. Finally, we also showed that certain environmental factors may drive the population size of specific yeast populations. The present results indicate that the wine yeast community has a geographical character at local scale, which is an important feature of the microbial terroir concept and thus for the wine industry.

Truth in wine yeast.

SummaryEvolutionary history and early association with anthropogenic environments have made Saccharomyces cerevisiae the quintessential wine yeast. This species typically dominates any spontaneous wine fermentation and, until recently, virtually all commercially available wine starters belonged to this species. The Crabtree effect, and the ability to grow under fully anaerobic conditions, contribute decisively to their dominance in this environment. But not all strains of Saccharomyces cerevisiae are equally suitable as starter cultures. In this article, we review the physiological and genetic characteristics of S. cerevisiae wine strains, as well as the biotic and abiotic factors that have shaped them through evolution. Limited genetic diversity of this group of yeasts could be a constraint to solving the new challenges of oenology. However, research in this field has for many years been providing tools to increase this diversity, from genetic engineering and classical genetic tools to the inclusion of other yeast species in the catalogues of wine yeasts. On occasion, these less conventional species may contribute to the generation of interspecific hybrids with S. cerevisiae. Thus, our knowledge about wine strains of S. cerevisiae and other wine yeasts is constantly expanding. Over the last decades, wine yeast research has been a pillar for the modernisation of oenology, and we can be confident that yeast biotechnology will keep contributing to solving any challenges, such as climate change, that we may face in the future.

ველური ვაზის სახელწოდებები საქართველოში და მისი ეთნობოტანიკური გამოყენება

Wild grapevine Vitis vinifera subsp. sylvestris (C.C.Gmel.) Hegi is a typical plant of Georgian flora, spread in our country. Consequently, a numerous number of local synonym names of this plant are available in different historical-ethnographic provinces of Georgia. Among these names ‘Krikina’, ‘Babilo’, ‘Mortskhula’, ‘Brdzghuamli’ and ‘Dzghvamli’ are more spread, having different spelling variations. The great contrubution of wild grape is that it, as an ancestor of cultivated grapevine, made the basis of grape breeding in Georgia and is a distant relative of our varieties. Besides, as it was demonstrated by the ethnobotanical research, due to long period present in the forests of Georgia the local people used this plant as a food (grape, pickles), a beverage (young fermented wine ‘Machari’, wine, grappa), a construction material (buildings, door for a church, breedge), a tool for washing Qvevri, a pollinator for grape cultivars, a honey plant, a rootstock, a planting material for vineyards.

Comparative Analysis of Glycosidic Aroma Compound Profiling in Three Vitis vinifera Varieties by Using Ultra-High-Performance Liquid Chromatography Quadrupole-Time-of-Flight Mass Spectrometry.

Glycosidic aroma compounds are the important precursors of volatile aroma in grapes, and they can be added with odorous aglycones via enzyme- or acid-catalyzed hydrolysis during wine fermentation and storage. Developing an analytical method for intact glycosides can provide the possibility to study the accumulation of these aroma precursors in grape berries. For this purpose, a Tandem Mass Spectrometry (MS/MS). database based on ultra-high-performance liquid chromatography quadrupole-time-of-flight mass spectrometry was built, covering multiple aglycone classes. Subsequently, the profiles of glycosidic aroma compounds in Vitis vinifera L. cv. Muscat Blanc, Riesling, and Chardonnay berries during maturation were investigated. Pentosyl-hexosides were the most abundant glycosides in all three varieties. Both composition and concentration of glycosidic aroma compounds varied obviously among grape varieties. Except for monoterpenol pentosyl-hexosides, most glycosides were kept almost stable in their concentrations during berry maturation. This research provides an approach to understand the variation of glycosidic aroma components from the perspective of aglycones and glycosides.

Using untargeted metabolomics to profile the changes in roselle (Hibiscus sabdariffa L.) anthocyanins during wine fermentation

In this study, an UHPLC-QE-MS approach in combination with multivariate statistical analyses was used to investigate the metabolic profiles, especially the anthocyanin profiles, during the fermentation of roselle wine. Overall, a large number of different metabolites (e.g., phenols, lipids, carbohydrates, amino acids and peptides, and others) were identified in the fermentation processes. Eight anthocyanin metabolites were identified in roselle wine for the first time, of which six were identified in the main fermentation process and two in the post-fermentation process. In addition, we identified several metabolic pathways during the fermentation process, and the metabolic pathways of anthocyanins in roselle wine are mainly related to flavonoid biosynthesis and to anthocyanin biosynthesis. These findings are expected to be useful for further studies on wine chemistry and yeast metabolism.

Microbial dynamics in industrial-scale wine fermentation employing Hanseniaspora uvarum β-glucosidase-producer strain.

The use of non-Saccharomyces yeast strains in winemaking is becoming a common trend. In fact, consumers are demanding new and healthier styles of wine. On the other hand, these strains are a challenge for the starting process due to winery-resident strains, especially with regard to industrial-scale fermentations. Current assay focuses on the scale-up of the laboratorial inoculum inside the winery environment to ferment 15,000 and 25,000 L of Vitis labrusca Bordô must, using a Hanseniaspora uvarum β-glucosidase-producer strain as starter culture. This scale-up could confirm the viability of using non-Saccharomyces yeast, as it presented promising results on a laboratory scale. The non-Saccharomyces strain was selected in a previous study since it proved to increase resveratrol concentration in lab scale winemaking. The yeast diversity was followed by the plate culturing method. Species identification and strain typing were determined by ITS-RFLP and PCR-fingerprinting, respectively. Physical and chemical analyses and resveratrol quantification were performed in the elaborated wines.

Development of a post-processing method to reduce the unique off-flavor of Allomyrina dichotoma: Yeast fermentation

Allomyrina dichotoma larva is an edible insect that has been consumed in East Asia for centuries. A. dichotoma is suitable for insect farming due to its numerous health benefits and advantages in industrial production. However, its market share is very poor due to their unique off-flavor, which negatively affects consumer preferences. So far, only one study has reported an effective method for reducing the unique off-flavor of A. dichotoma larvae. In this study, to reduce the off-flavor without increasing larval mortality, we inoculated Saccharomyces cerevisiae W-3, which is a wine fermentation yeast strain, in processed larva powder and investigated changes in volatiles. In the fermented larva powder, the intensity of indole, which has a strong fecal odor, decreased to 1/12. In addition, fruit-flavored volatiles such as ethyl acetate, isopentyl acetate, and 2-butanone increased by 17, 13, and 4 times, respectively. To provide clues for understanding the mechanisms by which volatiles change, we performed network analyses. Our yeast fermentation method can be used as a post process to reduce the off-flavor and improve the overall flavor of A. dichotoma larvae.