Traditional Alcoholic Beverage Research Articles

Comparative study between the effects of aged and fresh Chinese baijiu on gut microbiota and host metabolism

Aging is an essential part for production of the most distilled spirits. Aged spirits smell and taste more harmonious and commonly have higher antioxidant capacity in vitro than fresh spirits. However, it is unclear how aged spirits influence intestinal microbiota and host metabolism, which play important role in the development and progression of alcoholic liver disease (ALD). Baijiu is a traditional Chinese alcoholic beverage. Freshly distilled baijiu needs to be aged for at least 1 year. Here, we built an animal model through oral gavage by fresh baijiu, 10-year-old baijiu, 20-year-old baijiu, ethanol and water. Our results showed that mice treated with aged baijiu had lower degrees of liver injury than that of fresh baijiu as revealed by the phenotypic, biochemical and histologic analyses. In addition, significant different gut microbiota structures were observed between mice treated with aged baijiu and fresh baijiu. Further co-occurrence network showed that the aged baijiu groups had more complex microbial interaction than fresh baijiu group. By metabolomics, we observed aged baijiu and fresh baijiu intervention resulted in distinct serum metabolome. Sixty-four metabolites showed significant alteration among different interventions. Further metabolic pathway analysis based on these differential metabolites revealed 7 pathways were significantly impacted. A total of 13 metabolites were involved in these pathways, including 8 amino acids, 4 organic acids and 1 carbohydrate. Our results filled the gap of how aged spirits affect gut microbiota and host metabolism and provided supporting evidence that the aged spirits may have less effect on liver than fresh spirits.

Synthesis, Characterization, and Remedial Action of Biogenic p-Ag Nanoparticles

In the present study, the silver (Ag) nanoparticles (NPs) were fabricated using pakhoi (p), a traditional alcoholic beverage popularly used in the Garhwal region of Uttarakhand that has been known to possess significant antimicrobial activity properties. Different physicochemical techniques were used to characterize p-Ag NPs. The results confirm the synthesis of crystalline p-Ag NPs having a nearly spherical shape with a net positive charge. Further, p-Ag NPs exhibit strong antibacterial activity against Gram -ve bacteria. Moreover, a detailed study will be beneficial to understanding and exploiting the biomedical application and environmental remediation activity of the p-Ag NPs.

Registration of Geographic Indications of Alcohol Beverages from Abroad in Indonesia

The research aimed to describe why alcoholic beverage products belonging to foreign countries can be registered in Indonesia's Geographical Indications (GI) protection regime. It outlined the legal implications of protecting foreign alcoholic beverage products in the GI protection regime. The research was normative legal research which was strengthened by interviews. The research shows that the protection of foreign alcoholic beverages in the national GI regime is caused by two factors, including disharmony between law 20/2016 and Permenkumham number 12 the Year 2019, and the GI regime gives each member country the freedom to apply the GI provisions in accordance with their national interests. Furthermore, the implications for the protection of foreign alcoholic beverage products in the GI protection regime in Indonesia, besides showing the inconsistency of GI protection arrangements for alcoholic beverage products, also injure the sense of justice for traditional alcoholic beverage activists who want their traditional knowledge products to be protected through the GI regime.

Metagenomics analyses of microbial dynamics associated with putative flavor development in mash fermentation of sake

Sake is a traditional alcoholic beverage in Japan. To examine the impact of co-fermentation on microbial composition, fermentation with and without pineapple was conducted. The dynamics of microbial communities during fermentation were investigated using the metagenomics of 16S rRNA and internal transcribed spacers. In regular (RS) and co-fermented (CS) sakes, the fungal microbiota was dominated by Saccharomyces. Hierarchical clustering and non-metric multidimensional scaling identified two phases of bacterial succession. Compared with the dividing point between phases in RS, the transition in CS was delayed by five days. In both experiments, Leuconostoc dominated the premature stage and was replaced by Erwinia in the maturation stage; the alcohol tolerators of Staphylococcus and Bacillus were persistent. In RS, bacterial succession was characterized by a dramatic decrease in Pseudomonas in the maturation stage, whereas it occurred earlier in the premature stage in CS. Nineteen genera were exclusively found in CS, suggesting that co-fermentation may nurture diverse bacteria that enrich the sake flavor. Analysis of functional pathways by PICRUSt2 revealed that sugar alcohol degradation was fundamental to fermentation, whereas flavor-rich CS was attributable to the activation of l-arginine biosynthesis and amine and polyamine degradation. Aerococcus was positively correlated with enzymes involved in phenylethyl metabolism.

Stachydrine derived from fermented rice prevents diet-induced obesity by regulating adipsin and endoplasmic reticulum homeostasis

Makgeolli, a widely consumed traditional alcoholic beverage in Korea, is brewed mainly from rice using Nuruk as a fermentation starter, which contains fungi, yeast, and lactic acid bacteria. Among 58 Makgeolli samples brewed using various Nuruks, we found that one exhibited anti-obesity properties, with stachydrine shown to be responsible for these properties. Stachydrine promotes lipolysis and inhibits lipid accumulation in 3T3-L1 adipocytes; it also reduces weight gain and improves glucose tolerance and insulin resistance in a mouse model. Stachydrine dramatically suppresses adipsin mRNA levels in liver and adipose tissue, whereas serum adipsin levels were elevated in stachydrine-treated mice compared to mice fed a high-fat diet alone. Moreover, stachydrine recovers endoplasmic reticulum homeostasis and regulates adipsin expression. We highlight the potential use of stachydrine as a therapeutic agent for the treatment of obesity and insulin resistance and the use of Makgeolli fermented by Nuruk as a source of novel bioactive compounds.

Functional Characterization of khadi Yeasts Isolates for Selection of Starter Cultures.

Yeasts play an important role in spontaneous fermentation of traditional alcoholic beverages. Our previous study revealed that a mixed-consortia of both Saccharomyces and non-Saccharomyces yeasts were responsible for fermentation of khadi, a popular, non-standardized traditional beverage with an immense potential for commercialization in Botswana. Functional characterization of isolated fermenting yeasts from mixed consortia is an indispensable step towards the selection of potential starter cultures for commercialization of khadi. In this study, we report the characterization of 13 khadi isolates for the presence of brewing-relevant phenotypes such as their fermentative capacity, ability to utilize a range of carbon sources and their ability to withstand brewing-associated stresses, as a principal step towards selection of starter cultures. Khadi isolates such as Saccharomyces cerevisiae, Saccharomycodes ludwigii and Candida ethanolica showed good brewing credentials but Lachancea fermentati emerged as the isolate with the best brewing attributes with a potential as a starter culture. However, we were then prompted to investigate the potential of L. fermentati to influence the fruity aromatic flavor, characteristic of khadi. The aroma components of 18 khadi samples were extracted using headspace solid phase micro-extraction (HSSPME) and identified using a GC-MS. We detected esters as the majority of volatile compounds in khadi, typical of the aromatic signature of both khadi and L. fermentati associated fermentations. This work shows that L. fermentati has potential for commercial production of khadi.

Comprehensive two-dimensional gas chromatography mass spectrometry-based untargeted metabolomics to clarify the dynamic variations in the volatile composition of Huangjiu of different ages.

Aging plays an important role in the formation of aroma characteristics of Huangjiu, a traditional Chinese alcoholic beverage. Comprehensive two-dimensional gas chromatography mass spectrometry (GC×GC-qMS)-based untargeted metabolomics combined with a multivariate analysis was used to investigate the dynamic variations in the aroma profile of Huangjiu during aging process and to establish the relationship between the changing volatile metabolite profiles and the age-dependent sensory attributes. A total of 144 volatile metabolites were identified by GC×GC-qMS and 63 were selected as critical metabolites based on variable importance in projection values and p-values. Based on the results of principal component analysis, orthogonal partial least-squares discriminant analysis, and hierarchical clustering analysis, the samples of six different ages were divided into three groups: 1Y and 3Y samples, 5Y and 8Y samples, and 10Y and 15Y samples. The partial least-squares analysis results further revealed the relationship between the aromas attributes and variations of these volatile compounds. The high esters, aldehydes, and lactones contents contributed to the high intensities of the sweet and ester aroma attributes of the aged Huangjiu, while the high alcohols and ethyl esters contents contributed to the alcoholic and fruity aroma attributes of the newly brewed Huangjiu. These results improve our understanding of the chemical nature of the aroma characteristics of aged Huangjiu. PRACTICAL APPLICATION: Huangjiu is often labeled with its age as a measure of quality, which influences consumers' choice. Dynamic variations in volatile compounds of Huangjiu during aging and its contribution to the aroma characteristics of Huangjiu were figured out, which will assist the industry to produce better quality aged Huangjiu for consumers.

Wild and cultivated plants used in traditional alcoholic beverages in Italy: an ethnobotanical review

Wild and cultivated plants used in traditional alcoholic beverages in Italy: an ethnobotanical review

Inorganic fertilization improves Agave potatorum Zucc growth and nutrition

Agave potatorum Zucc is a wild agave used for mezcal (a traditional alcoholic beverage) production in Oaxaca, Mexico, but little information is available on its response to nitrogen (N) and phosphorus (P) fertilization. The effect of different N (0, 23.7, 47.5, and 71.2 mg N per kg soil) and P (0, 14.1, 28.3, and 42.5 mg P per kg soil) doses on A. potatorum growth, nutrition and total soluble solid in stem (TSS) was assessed using a two-factor completely randomized design under nursery conditions. Relative to the control, the highest increase in response variables was unfolded leaf number (UL) 37.3%, leaf relative chlorophyll (LRCC) 9.6% and leaf nitrate content (LNO) 42.7% with 71.2 mg N per kg soil; plant height (PH) 39.1%, stem diameter (SD) 17.2%, and leaf dry weight (LDW) 70.2% with 47.5 mg N per kg soil; leaf P content (LP) 78.2% with 23.7 mg N per kg soil; root density (RD) 28.5% and LRCC 5.5% with 28.3 mg P per kg soil; LP 34.7%, LNO 16.8% with 14.1 mg P per kg soil; and TSS 19.0% with 42.5 mg P per kg soil. The combined application of N and P also enhanced stem dry weight, root dry weight, root length, SD, LDW, PH, UL, LP, TSS, LNO and LCRR. The application of N and P favors growth, nutrition and TSS in A. potatorum; therefore, it could be an important alternative for agronomic management of this agave species under nursery conditions. Further research is recommended to assess the response of A. potatorum to N and P fertilization under field conditions.

Utilization of Sake lees as Broiler Feedstuff and its Effects on Growth Performance and Intestinal Immunity.

Increasing food loss and waste (FLW) is a global problem, and efforts are being made to use waste food as potential livestock feed material. The amount of self-supplied feed is lower in Japan than in other countries, and the government recommends FLW use for animal feed. Sake (Japanese rice wine) is a traditional alcoholic beverage. During the sake manufacturing process, large amounts of squeezed solids or “lees” (sake lees) are generated. Sake lees are nutritious and functional, but are prone to spoilage. In this study, we investigated whether sake lees should be mixed with animal feed immediately or after drying. To assess the usefulness of sake lees as a poultry feed ingredient and determine the effect of sake lees on intestinal immunity, we performed a feeding trial with three treatments: a raw sake lees (RSL) diet, dried sake lees (DSL) diet, and control diet. Three-week-old broilers were fed these diets (n=8 per group) for two weeks. We then calculated feed efficiency and performed RT-qPCR to assess the effects of diet on intestinal immunity. The growth performance in the RSL diet group was equivalent to that in the control diet group. The DSL diet became difficult for broilers to eat, resulting in decreased growth performance. In the ileum of RSL-diet broilers, the mRNA expression levels of TGF-β1 and avian β-defensin (AvBD)12 were significantly increased compared to those of control diet broilers (p<0.05), and a significant correlation was observed between the two genes (p<0.05). Our results indicated that sake lees should not be dried and should be mixed immediately with feed, and this sake lees when fed to chicken activates the intestinal immunity. However, sake lees have a lower fat content than corn, and it is thus important to combine sake lees with high-energy feed.

Crosstalk between gut microbiota and host lipid metabolism in a mouse model of alcoholic liver injury by chronic baijiu or ethanol feeding.

A growing body of evidence highlights the important role of gut microbiota and host metabolism, particularly for lipid metabolism, in the development and progression of alcoholic liver disease (ALD). However, the effects of fermented alcoholic beverages on gut microbiota and host lipid metabolism remain under-investigated. Moreover, the crosstalk between gut microbiota and host lipid metabolism is still unclear in experimental ALD. Baijiu is a traditional Chinese alcoholic beverage. It contains large amounts of small molecule bioactive compounds in addition to a significant amount of ethanol (EtOH). In this study, we showed that baijiu caused lower degrees of liver injury than pure EtOH as revealed by phenotypic, biochemical and histologic analyses. Furthermore, baijiu and EtOH gavage resulted in different gut microbiota structures. Specifically, the baijiu group had a significantly higher abundance of Ruminococcus, Oscillospira, Mucispirillum, Bilophila, Parabacteroides and Odoribacter and a lower abundance of Helicobacter and Prevotella than that of the EtOH group. Using a targeted metabolomics approach, we also observed a greater than 19% increase of total hepatic free fatty acids (FFAs) after baijiu feeding and a 33% increase of hepatic FFAs after EtOH feeding. Baijiu feeding significantly increased total hepatic mono-unsaturated FAs (MUFAs). In contrast, polyunsaturated fatty acids (PUFAs), MUFAs and saturated FAs were significantly increased by EtOH feeding. Finally, Spearman's rank correlation showed that the increased levels of FFAs (mainly C20 and C22 unsaturated FAs) significantly correlated with key different gut microbiota, including a positive correlation with Desulfovibrio, Maihella, Helicobacter, Acholeplasma, Parasutterella, Prevotella, AF12 and Alistipes, and a negative correlation with Dorea, Olsenella, Adlercreutzia and Akkermansia. Our results suggest that compounds in baijiu attenuated the development of ALD and thus provided supporting evidence that the host-gut microbiota metabolic interactions play an important role in the development of ALD.

How to Habits of Drinking Alcohol "SOPI" in Community?- Dental Caries Status

Dental and oral health problems in the people of East Nusa Tenggara Province are in the high category. The people of Coal Village have a habit of drinking sopi, a traditional alcoholic beverage. Frequently drinking alcoholic beverages for long duration has a negative risk to the oral health. Sopi is one of the traditional alcoholic beverages, fermented from Enau (palm) tree plants. The purpose of this research is relationship between the habits of drinking sopi and the status of dental caries. Method: This is a research of Analytical Survey. This research was conducted in November to December 2019. The population on this research was the society of sopi drinkers. The total sampling was 76 people. The data collecting of the sopi drinking habits was carried out by completing the checklists; the data of the dental caries status was obtained by direct examinations. The data analysis used the test of Kendall’s Tau c. Results: The duration of drinking sopi was classified in the old category (93,4%); the frequency of drinking sopi was classified in the frequent category (63,2%); the volume of the drunk sopi was classified in the lots category (61,8%); the habits of drinking sopi was classified in the heavy category (93.4%); and the dental caries status was classified in the high criterion (52,6%). Based on the correlation test results, that there was no relationship between the time duration, frequency, volume, and the habits of drinking sopi with the status of dental caries (P &lt; 0,05). Conclusion: There was no relationship between the habits of drinking sopi and the status of dental caries.

Isolation and analysis of a sake yeast mutant with phenylalanine accumulation.

Sake is a traditional Japanese alcoholic beverage brewed by the yeast Saccharomyces cerevisiae. Since the consumption and connoisseurship of sake has spread around the world, the development of new sake yeast strains to meet the demand for unique sakes has been promoted. Phenylalanine is an essential amino acid that is used to produce proteins and important signaling molecules involved in feelings of pleasure. In addition, phenylalanine is a precursor of 2-phenylethanol, a high-value aromatic alcohol with a rose-like flavor. As such, adjusting the quantitative balance between phenylalanine and 2-phenylethanol may introduce value-added qualities to sake. Here, we isolated a sake yeast mutant (strain K9-F39) with phenylalanine accumulation and found a missense mutation on the ARO80 gene encoding the His309Gln variant of the transcriptional activator Aro80p involved in the biosynthesis of 2-phenylethanol from phenylalanine. We speculated that mutation of ARO80 would decrease transcriptional activity and suppress the phenylalanine catabolism, resulting in an increase of intracellular phenylalanine. Indeed, sake brewed with strain K9-F39 contained 60% increase in phenylalanine, but only 10% less 2-phenylethanol than sake brewed with the parent strain. Use of the ARO80 mutant in sake brewing may be promising for the production of distinctive new sake varieties.One-Sentence SummaryThe ARO80 mutant is appropriate for controlling the content of phenylalanine and 2-phenylethanol.

Documentation of traditional alcoholic beverages and their indigenous utilization pattern by Kinnaura tribes of Himachal Pradesh, North Western Himalaya

Traditional alcoholic beverages are part and parcel of indigenous communities worldwide since ancient ages. These beverages are very common and popular among the indigenous communities residing in rural and tribal areas of India. In view of this, The present study has been conducted to document the traditional alcoholic beverages and their indigenous uses by Kinnaura tribes of Himachal Pradesh. Kinnaura, the tribal communities of Kinnaur district, Himachal Pradesh are famous for their own unique traditions, culture, history and lifestyle. The usage of local alcoholic beverages especially Angoori, Moori, Rashi, Brandi, Shudang, Chhang and Zanashang are quite common among Kinnaura tribes. These are important part of their customary rituals, festivals, diet and health since time immemorial and prepared by the tribal communities in every village across the Kinnauar district for their own use by following indigenous methods using fruits, grains and bulbs of 15 locally available species. The information on collection time, processing and production was gathered through interviews. Production of these traditional beverages are limited to household level and the methods involved for their preparation are almost similar all across the Kinnaur district. These indigenous practices are also being followed by the present generations through the knowledge gained from their elders. Legal permission for mass production, branding and popularization of traditional beverages and adoption as livelihood option has been suggested.

Ethnic preparation of Chubitchi, an alcoholic beverage of the Garo tribe of Meghalaya: a sociocultural analysis

The Garo community living in Meghalaya has always relied on fermented rice liquor known as Chubitchi as a main beverage in their everyday life. Chubitchi is celebrated by the Garos during feasts and ceremonies. The indigenous populations of North-East India prepare alcoholic beverage in their traditional ways, almost always using rice as the base for those drinks. Chubitchi is a popular traditional alcoholic beverage prepared by the Garo tribe of Meghalaya, India. It is consumed during festive occasions such as Wangala (harvest festival), Do.si Do.doka (wedding festivities) and religious rituals, and also served as refreshments for weary field labourers and guests. It has a great significance in the social and cultural life of the Garos. This paper deals with the traditional preparation of Chubitchi by the Garo tribe, which is a vital part of their culture, and also examines the sociocultural significance of the beverage.

Physicochemical properties, antioxidant activities and microbial communities of Ethiopian honey wine, Tej

Ethiopian honey wine, Tej, is spontaneously fermented traditional alcoholic beverage, usually made from honey and “gesho” (Rhamnus prinoides). Till now, limited amount of information is available on the characterization of Tej. Thus, the aim of this paper is to reveal the microbiological diversity and physicochemical properties of Tej samples collected from different areas of Ethiopia. High-throughput sequencing, electrochemical and chromatographic techniques, and spectrophotometric methods were used to achieve these objectives. Although there was a statistical difference in the exact values of physicochemical properties between the collected Tej samples, the pH and titratable acidity values of the samples ranged from 2.8 to 3.8 and from 1.81 to 8.65 g/L, respectively. Similarly, the alcohol and sugar contents of the samples were in the range of 6.36–11.34 g/100 mL and from 0.37 to 31.6 g/L, respectively. Moreover, the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 3-ethylbenzothiazoline-6-sulfonic acid diammonium salt (ABTS) values of the samples were in the range of 37.9–81.0% and 27.4–73.1%, respectively. Furthermore, microbial community structure was predominated by a few fermentative microorganisms. Specifically, the bacterial community structure was dominated by the genera of Lactobacillus (53.15%) and Zymomonas (38.41%). Whereas, the fungal community structure was exclusively dominated by genus of Saccharomyces (99.66%). Additionally, Lactobacillus, Zymomonas and Saccharomyces were the detected core microbiome for the collected Tej samples. Both bacterial and fungal communities had shown no statistically significant differences in alpha diversity analysis based on the area of sample collection. However, the bacterial communities had a statically significant difference in Unweighted Unifrac beta diversity analysis. Generally, the observed shared physicochemical characteristic features and the dominance by certain group of microorganisms might be seen as a boon for the development of direct fermentation system to this traditional alcoholic beverage.

Medicinal Plants in Traditional Herbal Wines and Liquors in the East of Spain and the Balearic Islands.

Homemade herbal preparations from the East of Spain are the witness of traditional medicine inherited from the ancient complex formulas of herbal teas and medicinal wines. In this study, we document the use of traditional alcoholic beverages, identify their ingredients, almost exclusively botanical, record the local medicinal uses of these mixtures, and discuss patterns of distribution of this knowledge in regions of eastern Spain, the Balearic Islands and Andorra. We determine marker species and relevant patterns of herbal formulas in the different regions of the territory. Homemade liquors and liqueurs are consumed for their digestive and tonic-restorative properties but they also play in some cases an important social role. The elderly remember other medicinal uses such as aperitif, emmenagogue, or antidiarrheal, for some of the most popular preparations. The herbal liqueur formulas include predominantly Lamiaceae, Asteraceae, Rosaceae, Rutaceae, and Apiaceae species. Herbs (58%), fruits (28%), and mixtures of both (12%) are ingredients of liquors and wines, being the aerial parts the most frequent in terms of species (30%) and records (49%). Dictamnus hispanicus, Santolina villosa, Salvia blancoana subsp. mariolensis, Rosmarinus officinalis, Thymus vulgaris, and Clinopodium serpyllifolium subsp. fruticosum are the species most frequently used. Others species used to a lesser extent as Polygonatum odoratum, Thymus moroderi, and Saxifraga longifolia are restricted to locally homemade preparations because their collection and uses require special knowledge of the rare or endemic flora. Sustainability of these practices is strongly limited by the overall loss of local traditional knowledge and by the limited availability of most of the wild species; some of them are endangered or threatened mainly by the loss of their natural habitats. Cultivation and domestication are a promising alternative to collecting from wild populations. The cultivation of Thymus moroderi in the province of Alicante and Polygonatum odoratum in the province of Teruel are good examples. There is a notable decrease in the complexity of the formulas registered throughout the nearly 15 years of the study. This is interpreted as a consequence of a loss of knowledge, less accessibility to wild resources, and changes in traditions and preferences.

Natural preservative‐based shelf‐life enhancement of borde: A traditional Ethiopian low alcoholic beverage

Borde is a cereal-based low alcoholic fermented beverage with a short shelf-life (fast spoilage due to over-acidification). Thus, the research aims to enhance the shelf-life. The counts of aerobic mesophilic bacteria and yeast in samples with 0.1 g/100 ml sodium benzoate and control sample (without preservatives) decreased considerably during the 14 days of storage. There were no significant differences (p > .05) in counts of lactic acid bacteria for 14 days in borde samples treated with all preservatives. Likewise, the counts of yeast showed no significant differences for 12 and 10 days between borde samples treated with 10 g/100 ml Moringa oleifera (6.48 ± 0.04 to 6.42 ± 0.01), and 10 g/100 ml Thymus schimperi (6.49 ± 0.03 to 6.00 ± 0.07), respectively. Borde samples treated with 10 g/100 ml T. schimperi were more acceptable than those treated with 10 g/100 ml M. oleifera. Hence, T. schimperi was the candidate preservative for the preservation of borde. Practical applications The practical application of the use of these natural preservatives is the feasibility and ease of the method for preservation of low alcoholic cereal-based beverages (borde) even at household level among those who cannot afford alternative preservatives. Besides, natural preservatives (Moringa oleifera and Thymus schimperi) are naturally safe, accessible, and nutritious in addition to their role in enhancing the shelf life of beverages. Thus, the current findings could pave way for further evaluation of these plants and others for their application in many other fermented products, be it alcoholic or non-alcoholic beverages.

Barriers to the Implementation of Alcohol Policies in Jayapura City

Background: Basic Health Research (RISKESDAS) in 2018 found as many as 15 provinces out of 35 provinces had a prevalence of alcohol consumption above the national prevalence, and Papua was reportedly in the top 15 of the prevalence of high alcohol consumption (alcohol) as much as 4.6%. Objective: To describe the obstacles to the implementation of local government policies in curbing the circulation of alcohol in Jayapura City. Method: This is a descriptive study using a qualitative single instrumental case study design in Jayapura City from May to September 2020. Purposive sampling technique was employed to determine participants who consisted of an integrated team of alcohol control and supervision (n =6). Data collection was using in-depth interviews and non-participant observation. Result: Communication between the integrated team was not optimal. National Food and Drug Agency (BPOM) had not routinely conducted sample tests as the majority of traditional alcoholic beverages sold in shops such as local liquors were illegal. The division of duties and responsibilities by the integrated team was good, but the execution in the field was not as agreed upon. There were alcohol traders who have not applied for licenses to sell alcohol because the process to get license permits was too complicated. The content of Perda No.8 of 2014 weakened the police because they could not give punishment like Satpol PP, however, Satpol PP did not have an investigating team. Conclusion: The lack of commitment from the control and supervision team of alcohol circulation in carrying out their duties. The resources involved were lacking. Coordination among the integrated teams was not going well thus the division of tasks and authorities with implementation actions in the field did not work according to the policy enforce in Jayapura City. Evaluation of work programs by policy implementers could be carried out to increase the success of implementation.

Role of Gln79 in Feedback Inhibition of the Yeast γ-Glutamyl Kinase by Proline.

Awamori, the traditional distilled alcoholic beverage of Okinawa, Japan, is brewed with the yeast Saccharomyces cerevisiae. During the distillation process after the fermentation, enormous quantities of distillation residues containing yeast cells must be disposed of, and this has recently been recognized as a major problem both environmentally and economically. Proline, a multifunctional amino acid, has the highest water retention capacity among amino acids. Therefore, distillation residues with large amounts of proline could be useful in cosmetics. Here, we isolated a yeast mutant with high levels of intracellular proline and found a missense mutation (Gln79His) on the PRO1 gene encoding the γ-glutamyl kinase Pro1, a limiting enzyme in proline biosynthesis. The amino acid change of Gln79 to His in Pro1 resulted in desensitization to the proline-mediated feedback inhibition of GK activity, leading to the accumulation of proline in cells. Biochemical and in silico analyses showed that the amino acid residue at position 79 is involved in the stabilization of the proline binding pocket in Pro1 via a hydrogen-bonding network, which plays an important role in feedback inhibition. Our current study, therefore, proposed a possible mechanism underlying the feedback inhibition of γ-glutamyl kinase activity. This mechanism can be applied to construct proline-accumulating yeast strains to effectively utilize distillation residues.