Traditional Alcoholic Beverage Research Articles

Lentilactobacillus hilgardii H-50 strongly inhibits lipopolysaccharide-induced inflammatory responses in mouse splenocytes via its specific surface layer proteins.

Certain lactic acid bacteria (LAB) are known to have anti-inflammatory effects; however, hiochi bacteria, which are taxonomically classified as LAB and known to spoil a traditional Japanese alcoholic beverage, have not been studied in the same context. The aim of this study is to investigate the anti-inflammatory effects of hiochi bacteria strains and the underlying mechanisms. We screened 45 strains of hiochi bacteria for anti-inflammatory effects and found that Lentilactobacillus hilgardii H-50 strongly inhibits lipopolysaccharide (LPS)-induced secretion of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 in mouse splenocytes. This inhibition is attributed to its specific surface layer proteins (SLPs), which directly bind to LPS. The L. hilgardii H-50 strain exerts anti-inflammatory effects through its SLPs.

Production and evaluation of wine prepared from banana (Saccharomyces Cerevisiae)

The article presents review on potential of wine production from Banana. Traditional alcoholic beverage has become common because of economic issue. This work was aimed to improve production process of alcoholic beverage based banana extract and to evaluate sensory parameters of the obtained alcoholic beverage. Juice was extracted from banana (Musa sapientum) pulp with was inoculated with Baker's yeast (Saccharomyces Cerevisiae)and naturally grown mycelium and held at 30+2'c for 14 days. Banana, a wonderfully sweet fruit with firm and creamy flesh that come prepackaged in a yellow jacket, available for harvest throughout the year consists mainly of sugars and fibers which make it a source of immediate and slightly prolonged energy. When consumed, reduces depression, anemia, blood pressure, stroke risk, heartburns, ulcers, stress, constipation and diarrhea. The physical chemical parameters was determine during the fermentation using a standard procedures like pH , type of alcohol (Luca’s Test ) specific gravity, alcohol content, preliminary phytochemical screening and reducing sugar. Alcohol content of wine is increase with time at specific temperature as increase in sugar concentration.

Improvement of valine and isobutanol production in sake yeast by Ala31Thr substitution in the regulatory subunit of acetohydroxy acid synthase.

The fruit-like aroma of two valine-derived volatiles, isobutanol and isobutyl acetate, has great impact on the flavour and taste of alcoholic beverages, including sake, a traditional Japanese alcoholic beverage. With the growing worldwide interest in sake, breeding of yeast strains with intracellular valine accumulation is a promising approach to meet a demand for sakes with a variety of flavour and taste by increasing the valine-derived aromas. We here isolated a valine-accumulating sake yeast mutant (K7-V7) and identified a novel amino acid substitution, Ala31Thr, on Ilv6, a regulatory subunit for acetohydroxy acid synthase. Expression of the Ala31Thr variant Ilv6 conferred valine accumulation on the laboratory yeast cells, leading to increased isobutanol production. Additionally, enzymatic analysis revealed that Ala31Thr substitution in Ilv6 decreased sensitivity to feedback inhibition by valine. This study demonstrated for the first time that an N-terminal arm conserved in the regulatory subunit of fungal acetohydroxy acid synthase is involved in the allosteric regulation by valine. Moreover, sake brewed with strain K7-V7 contained 1.5-fold higher levels of isobutanol and isobutyl acetate than sake brewed with the parental strain. Our findings will contribute to the brewing of distinctive sakes and the development of yeast strains with increased production of valine-derived compounds.

Front Cover: Phase‐Separated Structures of Sake Flavors‐Containing Cell Model Membranes (Chem. Biodiversity 1/2023)

Graphical Abstract Front Cover. Sake (a traditional Japanese alcoholic beverage) contains flavors such as ethyl caproate (EC) and Isovaleraldehyde (IVA). EC, is known as apples or pineapples flavor, enhances its economic value of beverage. Some people enjoy IVA. After adding EC or IVA, it was observed that decreasing phase separations on cell-sized liposomes made from 1,2-dioleoyl-sn-glycero-3-phosphocholine, dipalmitoyl-sn-glycero-3-phosphocholine, and cholesterol. The findings of this study enhance not only our understanding of the biological function of flavors but also aid in rapid and cost effectively analysis, as reported by Tsuyoshi Yoda in his research article at 10.1002/cbdv.202200750.

Occurrence of bisphenols and contribution of edibles liquids conditioned in plastic packaging to the dietary exposure in Cameroon

Bisphenols are known to be endocrine disruptors. They are ubiquitous in the environment and also in plastic materials such as those used in food packaging. They can lead to the contamination of food in contact with environmental products or plastic packaging. In Africa, there a few studies on the assessment of the amount of bisphenols in foods; hence, most African countries do not have policies guiding on their utilization. The present work is aimed at assessing bisphenol levels in sachet water, bagged whisky, crude palm oil and traditional alcoholic beverages of Cameroon, in Central Africa. A sensitive method based on liquid chromatography tandem mass spectrometry (UPLC-MS/MS) was developed and validated for the simultaneous determination of bisphenol A, B and F (BPA, BPB and BPF). This method showed that recoveries were from 72 to 120% and the limits of quantification for BPA, BFB and BPF from aqueous samples were 0.09, 0.10, and 0.09 µg L−1 respectively. Estimation of the contribution to dietary exposure of BPA from these matrices were lower than temporary Tolerable Daily Intake (t-TDI) (4 µg/kg of body weight/day) defined by The European food safety authority (EFSA). One brand of sachet water contained BPF at 14.5 ± 1.1 µg L−1.

Enhancement of the Functional Properties of Mead Aged with Oak (Quercus) Chips at Different Toasting Levels.

Consumers increasingly prefer and seek functional beverages, which, given their characteristics, provide important bioactive compounds that help prevent and treat chronic diseases. Mead is a traditional fermented alcoholic beverage made from honey solution. The aging process of mead with oak chips is innovative and bestows functional characteristics to this beverage. Thus, in this study, we sought to develop and characterize a novel functional beverage by combining the health benefits of honey with the traditional aging process of alcoholic beverages in wood. Phenolic compounds, flavonoids, and antioxidant capacity were analyzed in mead using oak chips at different toasting levels and aged for 360 days. LC-ESI-QTOF-MS/MS was used to analyze the chemical profile of different meads. Over time, the aging process with oak chips showed a higher total phenolic and flavonoid content and antioxidant capacity. Eighteen compounds belonging to the classes of organic acids, phenolic acids, flavonoids, and tannins were identified in meads after 360 days. Our findings revealed that the addition of oak chips during aging contributed to p-coumaric, ellagic, abscisic, and chlorogenic acids, and naringenin, vanillin, and tiliroside significantly impacted the functional quality of mead.

Phase-Separated Structures of Sake Flavors-Containing Cell Model Membranes.

Sake (a traditional Japanese alcoholic beverage) contains ethyl caproate (EC), which enhances its economic value. Isovaleraldehyde (IVA) is also a well-known flavoring agent in alcoholic beverages, which some people enjoy. Recently, studies revealed that EC decreased the size of homogenous 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC) liposomes whereas IVA increased their size. Cholesterol (Chol) and ergosterol were previously referred to as animal and fungus sterols. For the first time, this study demonstrated the phase behavior of the membrane in cell-sized liposomes containing EC and IVA. After adding EC, the solid ordered/liquid disordered (Ld) and liquid ordered (Lo)/Ld phase separation in DOPC/dipalmitoyl-sn-glycero-3-phosphocholine/cholesterol or ergosterol ternary membranes decreased, but the Lo/Ld phase separation decreased after adding IVA. Biophysics, physiological, and application aspects of EC and IVA evaluation were discussed. The findings of this study not only enhance our understanding of the function of flavors but also provide rapid and cost effective performance for the measurement.

Edukasi pencegahan konsumsi alkohol pada remaja di SMK Muhammadiyah 3 Singosari Malang

Alcohol use in adolescents needs serious attention because of the negative impact on adolescent health and development. The prevalence of alcohol consumption among adolescents in Indonesia is 3.3% of them starting to drink at the age of 10, and most of them are traditional alcoholic beverages (38.7%). This community service program aims to reduce alcohol consumption among teenagers. There are two activities involved in this community service: education on alcohol prevention and screening of alcohol consumption. We carried the activity at Muhammadiyah 3 Singosari Vocational High School Malang. The participants were 57 students. This activity resulted in an increase in students' understanding of preventing alcohol consumption, with an average before the education of 5.40. In contrast, after being given education, it's score increased to 6.16. Most of the students at SMK do not consume alcohol or are shown in the normal category of 86.9%. However, some still consume 3.3% alcohol in the heavy category. This activity concludes that the provision of education can increase the understanding of adolescents, and then it is hoped that it will prevent alcohol consumption among adolescents. Follow-up efforts from the screening results are carried out by continuing to provide education, counseling, and further diagnostic examinations for adolescents.

Effect of different moulds on oenological properties and flavor characteristics in rice wine

Rice wine is one of traditional alcoholic beverages in China, which were fermented from rice with moulds and yeasts. In this study, in order to determine the difference in rice wine brewed by four moulds, four types of rice wine (named as HIM, BM, HEM, and GM) on physical and chemical factors, free amino acids (FAAs), volatile and non-volatile metabolites, and sensory evaluation were investigated. Results showed that 17 FAAs, 186 volatile compounds and 489 non-volatile metabolites were detected in rice wine. However, metabolites of rice wine brewed by moulds were quite different, in HIM rice wine, the content of FAAs (2861.62 ± 109.49 mg/L), alcohols (32.91 ± 3.37 mg/L), esters (19.34 ± 0.69 mg/L) and phenols (10.89 ± 0.27 mg/L) were the highest among rice wine samples. The number of different non-volatile metabolites in four rice wine samples were 27, 20, 62, and 20 (HIM, GM, HEM, and BM) by Linear Discriminant Analysis Effect Size respectively. And most of these non-volatile metabolites were organic acids (pyruvic acid, shikimic acid, etc.). Sensory evaluation showed that rice wine (HIM) brewed with Aspergillus oryzae were better than other rice wine samples.

Characterization of a new Saccharomyces cerevisiae isolated from banana stems and its mutant with l-leucine accumulation for awamori brewing.

We isolated a new strain of the yeast Saccharomyces cerevisiae, 35a14, from banana stems in Okinawa. This strain did not belong to any industrial yeast groups in a phylogenetic tree and produced high levels of alcohol. Furthermore, awamori, an Okinawa's traditional distilled alcoholic beverage, brewed with an l-leucine overproducing mutant derived from 35a14 showed a high concentration of isoamyl acetate.

Controversy and the Role of the Existence of the Traditional Alcohol Beverages (Arak Bali) Industry on the Development of Custom Village

Aims: The discussion of traditional alcoholic beverages is an interesting and sensitive subject. Arak Bali is one of the traditional Balinese alcoholic drinks. This study examines further the controversy over the Balinese fermented and/or distillated beverage industry, especially after various regulations have been issued. In addition, this research also needs to pay attention to the role of the arak industry in the development of traditional villages.
 Study Design: Descriptive study.
 Place and Duration of Study: This study held during in the middle of 2022.
 Methodology: This study is a descriptive study, where data were collected through FGDs, literature reviews, and questionnaires. The data were analyzed using descriptive analysis techniques, both qualitatively and descriptively.
 Results: Arak Bali as a prospective industry to improve the economic welfare of the community, by promoting it as a heritage-based product. The results of the FGD mapped several roles of the Arak Bali industry for the development of Traditional Villages, including community income, community empowerment, economic development of the Traditional Village, implementation of religious rituals, togetherness in the social community, preservation of local wisdom. The role of the existence of the Arak Bali industry in the Traditional Village which is the center of the industry is certainly greater in almost all indicators of traditional village development. Meanwhile, the role of the wine industry in religious ritual activities does not differ between traditional villages which are the center and non-centra of the Arak Bali industry.
 Conclusion: In addition to consumption, Arak Bali is also used in religious rituals and traditional medicine. In general, the role of the wine industry in traditional villages in Bali is divided into economic functions, religious functions, and social functions. Where in the traditional village center for the Arak Bali industry, the role of the economy (income and community economic empowerment) is the most prominent. Whereas in the traditional village centers and non-centra of the wine industry, the role in religious rituals, togetherness in social society and the preservation of local wisdom are important.

ANTIBACTERIAL EFFICACY OF PALM WINE AGAINST COMMON BACTERIAL ISOLATES FROM VARIOUS CLINICAL SAMPLES

Palm wine is a traditional alcoholic beverage produced by natural fermentation of the sap of palm trees. This drink is common in various parts of the Asia & Africa and it is known by various common names such as Maharashtra (Tadi, Shindhi). The unfermented sap is called neera (Palghar in Maharashtra) and is refrigerated, stored and distributed by semi-government agencies. This study was conducted to investigate the antibacterial potential of palm wine on as an alternative to antibiotics by studying its antibacterial efcacy again common bacterial isolates from clinical samples.Freshly top-tapped palm wine samples were obtained from a local palm wine tapper in Palghar, Maharashtra state, India. Various strains of Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae and Staphylococcus aureus was carried out using agar diffusion method. The palm wine used in this assay inhibited the growth of all the selected bacterial isolate used. The inhibition ranged from 2.3 mm- 29.5mm. Palm wine subjected to fermentation at room temperature for 7 days (168 hours). Growth inhibitory effect of undiluted palm wine superior than groups of antibiotics such as Ampicillin, Ampliclox, Chloramphenicol, Tetracycline and Ciprooxacin.This study established that palm wine possess considerable antibacterial activity against selected common bacterial isolates. Since palm wine has shown potent antibacterial activity on the test bacteria, further clinical trials will be required that will prove the potential of palm wine as an alternative to antibiotics treatment among patients suffering from common infection caused by there organism.

Effect of Giving Traditional Drinks Kameko and Pongasih from Southeast Sulawesi on Sperm Quality of Mice (Mus musculus).

<b>Background and Objective:</b> The traditional beverage of Southeast Sulawesi is Kameko and Pongasih, which is a traditional alcoholic beverage. One of the bad effects of alcohol on reproductive health is the occurrence of infertility. This research aimed to determine the effect of consuming traditional beverages of Kameko and Pongasih on the quality of sperm of mice (<i>Mus musculus</i>). <b>Materials and Methods:</b> This research used a true experiment. The samples were some healthy mice (<i>Mus musculus</i>) aged 6-8 weeks as much 24 tails and divided into 4 groups. On the 36th day after the treatment was completed, the mice were terminated and prepared for microscopic sperm quality analysis. The quality of sperm includes motility, viability, concentration and morphology. The data test uses the Shapiro-Wilk Test, One-way ANOVA and <i>post hoc</i> LSD Test. <b>Results:</b> Kameko traditional beverage had a significant influence on sperm quality with motility, morphology, sperm concentration (p = 0.000) and viability (p = 0.001). Pongasih traditional beverage has a significant influence on sperm quality with motility parameter, morphology, sperm concentration (p = 0.000) and viability (p = 0.001). Kameko and Pongasih traditional beverages did not have a significant difference in sperm quality with motility parameter (p = 0.463), viability (p = 1.000), morphology (p = 0.553) and amount of concentration (p = 0.714). <b>Conclusion:</b> South East Sulawesi traditional beverages Kameko and Pongasih have effects on mice's (<i>Mus musculus</i>) sperm quality, but there was no difference in the group of mice which have been given Kameko and Pongasih.

The effects of biogenic amines in Chinese Huangjiu on the behavior of mice and hangover headache-related indices.

Huangjiu (Chinese rice wine) is a popular and traditional alcoholic beverage in China; however, the consumption of Huangjiu readily results in hangover symptoms. The aim of this study was to identify the main components associated with behavioral inhibition, headache, and the relevant mechanisms by using a mice hangover model. The results of an open-field experiment revealed that the key biogenic amine associated with mice behavior was histamine, which inhibited the behavior activity of mice in a dose-dependent manner. Moreover, histamine treatment decreased the levels of serotonin (5-HT) and 5-hydroxyindole acetic acid. In addition, the levels of dopamine and nitric oxide, which are associated with migraine, increased in the brain tissue of mice. In addition, the expression of receptor genes of 5-HT, including Htr1a, Htr1f, and Htr2c, is essential in regulating various behaviors and mental activities. In conclusion, the present study demonstrated that histamine is a key component in Huangjiu, and it is related to hangover symptoms by affecting the level of 5-HT and its receptors.

Development of mixed starter culture for the fermentation of Ethiopian honey wine, Tej

Ethiopian honey wine is one of the country's most popular spontaneously fermented traditional alcoholic beverages. However, the final product of this natural fermentation system is frequently of poor and inconsistent quality. Furthermore, it makes the process difficult to predict, control, and correct. Thus, the main aim of this study was to develop a direct fermentation system for Ethiopian honey wine, Tej. After isolating fermentative microbial strains from Tej samples, they were subjected to intensive screening to fit to its purpose. Later, phenotypic and genotypic characterization, and inoculation of isolates to honey-must were performed sequentially. Finally, microbial interaction and physicochemical analysis, including volatile compounds profiling, were done for the inoculated samples. The identified isolates were strains of Saccharomycetaceae and Lactobacillaceae families. These strains showed a good ability to tolerate osmotic stress and a lower pH environment. Tej sample produced by mixed culture inoculation of Saccharomyces and Lactobacillus species showed similar physicochemical, volatile compounds, and sensory attributes values with that of the control sample. Thus, a mixture of Saccharomyces and Lactobacillus strains could be used as a starter culture to produce Ethiopian honey, Tej, without scarifying of its major quality attributes.

Safety evaluation and comparative genomics analysis of the industrial strain Aspergillus flavus SU-16 used for huangjiu brewing

Huangjiu is a popular Chinese traditional alcoholic beverage, while its brewing processes have rarely been explored. We herein report the first gapless, near-finished genome assembly of the industrial strain Aspergillus flavus SU-16 for huangjiu brewing. This work provides insights and supports for the further industrial applications of A. flavus isolates by comprehensively studying of the safety and genomic variations of SU-16. We demonstrated that SU-16 is a non-aflatoxigenic A. flavus at both molecular and metabolic levels. Using of nanopore sequencing technology resulted in a complete genome sequence for all 8 A. flavus chromosomes, as well as the mitochondrion. Genome comparisons of SU-16 with reference strains identified the chromosomal rearrangements, revealed the adaption mechanism of SU-16 to huangjiu ecological niche, and found that SU-16 is a good repository for CAZymes and some bioactive secondary metabolites. The results will help to develop more scientific huangjiu fermentation processes, and explore metabolism pathways of desired or harmful components in huangjiu to improve its quality.

Comparison of fungal communities and nonvolatile flavor components in black Huangjiu formed using different inoculation fermentation methods.

Black Huangjiu (BH) is a traditional alcoholic beverage in China, which is very popular among people. The different methods (simultaneous inoculation, sequential inoculation), were applied to ferment BH in this study, which were investigated the changes in the composition of fungal communities and non-volatile flavor compounds (nVFCs) by high-throughput sequencing (HTS) and ultraperformance liquid chromatography–tandem mass spectrometer (UPLC MS/MS). The results showed that Rhizopus and Saccharomyces were the predominant fungal genera throughout fermentation, and 471 nVFCs were detected in BH after fermentation. Compared to that observed simultaneous inoculation, Rhizopus increased at the end of sequential fermentation, and the contents of the organic acids and their derivatives increased significantly [variable importance in the projection (VIP) > 1.0, p < 0.05, fold change (FC) > 2], while that of lipids and lipid-like molecules decreased significantly (VIP > 1.0, p < 0.05, FC < 0.5). Through the correlation analysis of 32 nVFCs with significant differences (VIP > 1.0, p < 0.05, FC >32 or < 0.03) and the community, it was found that lipids and lipid-like molecules (12) and organic acids and their derivatives (10) were significantly (p < 0.05) negatively correlated with Saccharomyces, but they were significantly (p < 0.05) positively correlated with Rhizopus. Compared with simultaneous inoculation, BH fermented by sequential inoculation, the taste was stronger, sweeter, mellow, and softer. Our findings provide information on nVFC dynamics and will aid in the selection of beneficial strains to improve BH quality.

Valorization of Traditional Alcoholic Beverages: The Study of the Sicilian Amarena Wine during Bottle Aging

Traditional alcoholic beverages have always been part of the Mediterranean culture and, lately, they have been re-evaluated to valorize both the territory and local customs. In this study, the Amarena wine, a fortified wine included in the national list of the traditional agri-food products, was characterized during bottle aging for oenological parameters, chromaticity, volatiles, and inorganic elements. Then, experimental data were visually interpreted by a principal component analysis (PCA). PCA revealed that most of oenological parameters (i.e., alcoholic grade, total dry extract, sugars, organic acids, and phenolic compounds) had a scarce discriminating power. Additionally, ethyl esters were only present in younger products, while remaining at quite constant levels. Conversely, certain metals (i.e., Mg, Na, Mn, Zn, and Cu), chromatic properties, and pH differentiated older Amarena bottles from the younger counterpart. Particularly, acetaldehyde and furanic compounds proved to be valid aging markers. A sensorial analysis highlighted that fruity and floral odors and flavors characterized younger beverages, while dried fruity, nutty, and spicy notes were displayed by older products, along with the valuable attribute of “oxidized” typically observed in aged Sherry wines. Overall, this study may encourage the production and commercialization of the Amarena wine, thus preserving the cultural heritage of the Mediterranean area.

A review on the nutritional composition, phytochemicals, and health benefits of barberry: An insight into culinary applications and future prospects

Barberry is a spinous shrub native to the Northern areas of the Himalayas and is widely distributed in the tropical regions of Asia, South America, Africa, and some parts of Europe. The berries of this shrub are eaten as a dessert and are a good source of minerals, total phenols, and total flavonoids. The major minerals are zinc (11.2 mg 100 g−1), iron (180.8 mg 100 g‒1), magnesium (5.8 mg 100 g−1), and potassium (526.4 mg 100 g−1). The concentration of total phenols and total flavonoids is reported to be 50.44 mg GAE/100 g and 6.08 CE/100 g, respectively. The root and bark are rich in alkaloids. The major alkaloids are berberine, berbamine, palmatine, and jatrorrhizine, etc. These alkaloids along with phenolics and flavonoids possess antidiabetic, hepatoprotective, immunomodulatory, anticancerous, antimicrobial, anti-inflammatory, antioxidant, antidiarrhoeal, anxiolytic, and hypolipidemic properties. The higher doses of its major alkaloid “berberine” can cause many health implications. So, precautions must be taken before its consumption. This manuscript discusses in detail the various nutritional and phytochemical properties of the plant to have its commercial application in food and nutraceutical products development. Practical applications Barberry fruits are eaten as dessert and are also used for the preparation of wine. Roots of this plant are used in the preparation of some traditional alcoholic beverages and medicines. A very valuable Ayurvedic preparation “Rasaut” is made from the roots and is used in the treatment of skin diseases, menorrhagia, diarrhea, jaundice, and ophthalmic infections. The other parts of this plant are also rich in alkaloids and flavonoids that possess antidiabetic, hepatoprotective, immunomodulatory, anticancerous, antimicrobial, anti-inflammatory, antioxidant, antidiarrhoeal, anxiolytic, and hypolipidemic properties.

Bioethanol production from biodegradable wastes using native yeast isolates from Ethiopian traditional alcoholic beverages

Due to the existing energy crisis and scarcity of conventional supplies of fossil fuels, there is a strong desire for the production of bioethanol from sustainable biodegradable waste as an alternative energy source. The aim of this study was to produce bioethanol from grain waste flour (GWF), Tella spent (TS), pineapple (PAP) and papaya peels (PYP) using native yeasts isolated from Ethiopian traditional alcoholic beverages. Diluted H2SO4 was used to hydrolyze polysaccharides in the waste materials. Starch content from GWF and reducing sugar concentration from each hydrolysate were determined by the standard protocol. Seventy-five native yeasts were isolated from local beverages for ethanol production. The 2% H2SO4 pretreatment produced the highest reducing sugar of GWF (30.1%); PYP (28.6%); PAP (16.4%); and TS (10.4%). In addition to this, GWF had 60.6% starch content. Of 75 isolates, 12 were able to tolerate ≥8% ethanol concentration, and 2 isolates, Y10 and Y55 showed strong ethanol production capacity (glucose to ethanol conversion efficiency) with 8.2% (79.8%) and 9.6% (94.3%) from synthetic medium, respectively. The highest ethanol yields of 16.1%, 9.8%, 8.0% and 6.1% were produced from GWF, PAP, PYP and TS, respectively using isolate, Y55 (identified as S. cerevisiae) within 168 h for GWF, 120 h for PAP and 72 h for PYP and TS. This study revealed that native yeast isolates could be used for industrial production of bioethanol via cheap biodegradable wastes.