Sake Production Research Articles

Influence of Starch Characteristics on Digestibility of Steamed Rice Grains under Sake-making Conditions,and Rapid Estimation Methods of Digestibility by Physical Analysis

Retrogradation of starch in steamed rice grains under sake-making conditions was examined. The enthalpy change derived from retrogradation of amylopectin increased during the koji and sake mash processes. Ethanol in the sake mash accelerated retrogradation of amylopectin. Enzyme digestibility and properties of endosperm starches of 136 Japanese rice samples used for sake production were examined. A digestion test of steamed rice grains was conducted after 24 h storage at 15°C to estimate more accurately digestibility during the sake-making process. The ratio of short-to-long chain amylopectin exhibited a significant correlation with enzyme digestibility, suggesting that the structure of amylopectin may be used as a predictor of digestibility. The gelatinization temperature of brown/milled rice and purified starch measured by DSC (differential scanning calorimetry) correlated well with the ratio of short-to-long chain amylopectin and with enzyme digestibility. Furthermore, the pasting temperature of milled rice as measured by RVA (Rapid Visco Analyser) correlated well with the ratio of short-to-long chain amylopectin and with enzyme digestibility. Therefore, DSC and RVA appear to be useful tools for estimating enzyme digestibility of steamed rice grains under sake-processing conditions because the measurements are done rapidly and require only small weights of brown/milled rice flour samples.

The Unification of Trademarks and the Change of Sales Strategies by a Nada Brewer

This paper clarifies the structure of the management development in prewar days of a large-scale brewer in Nada, Hyogo Prefecture, observing the source of the profit in the distribution process. Tatsuuma-Honke in Nishinomiya is adopted as an example. The way that should be and the change of the complicated Japanese sake distribution process are considered by an analysis which combined its sales market, sales strategy, and individual management. As a result, the following points were solved in this paper.First, it is confirmed that in the mid-1920s Tatsuuma-Honke established even in Tokyo the relation of maker predominance which was previously being developed in a local market. Although the company did not necessarily persist in its share reservation in the Tokyo market, without being bound by the monopoly right of the brand which remained among each wholesaler, it was meaningful to enable sales with the “Hakushika” brand which it specified itself.In process of the trademark establishment, the production of self-made sake increased and the raising of the bottom level of the quality was achieved. As a result, in Tatsuuma-Honke, it became possible to use a sales strategy which was further conscious of the brand's bottling sales. However, it was more important for Tatsuuma-Honke to internalize the structure which combined a sales method — undiluted sake dealing in a tun unit— called “Oka-Uri” with the opposite character of bottling, and carries out the hedge of the unsold risk. This also meant the possibility that the purchase of the undiluted sake from a minor-scale brewer would be demoted to the 2nd buffer for a large-scale brewer. This indication adds new knowledge to a simple understanding that the former specialized in Oke-Uri and the latter was based on sales of brand sake, as shown by the conventional history of research.The high profitability in the distribution process as the above-mentioned result relatively stabilized the brewing management of Tatsuuma-Honke in the depression of the 1920s. Such a case was just a pattern of the development which preceded the large-scale management in the prewar-days term of the Nada brewing industry.

口かみ「もろみ」に添加した清酒酵母の消長

Sake yeast with the fermentation ability for glucose, maltose and maltotriose was added to Kuchikami Rice Moromi. Increases and decreases of the microorganisms, the general composition, and the composition of saccharide were investigated, and the influence on the alcoholic production of the added sake yeast in Kuchikami Rice Moromi was examined. Though the added sake yeast increased in the early period of Kuchikami Rice Moromi, it decreased and disappeared about 5 days later. After that, though Saccharomyces sp.(wild type) appeared and increased, the number of cells was 106/ml. After the added sake yeast disappeared, the fermentation continued and the alcoholic production increased. This phenomenon was due to the alcoholic fermentation by Sacch. sp.(wild type), and was not influenced by the added sake yeast. Sacch. sp. and Lactic acid bacteria existed universally in Kuchikami Rice Moromi. The increase and decrease of those microorganisms was not constant as is Kimoto in sake production.

Structural and Retrogradation Properties of Rice Endosperm Starch Affect Enzyme Digestibility of Steamed Milled‐Rice Grains Used in Sake Production

ABSTRACTStructural and physicochemical characteristics of endosperm starch from milled rice grains of seven Japanese cultivars used in sake production were examined. Amylose content was 15.2–20.2%, number‐average degree of polymerization (DPn) of amylose was 900–1,400, and the ratio of short‐to‐long chain amylopectin was 2.7–3.5, respectively. The degree of retrogradation of purified starch stored for seven days at 4°C after gelatinization was 20–31%. The degree of retrogradation correlated negatively with the ratio of short‐to‐long chain amylopectin. The effect of holding time after steaming on enzyme digestibility and starch retrogradation of steamed rice grains was investigated. The longer the holding time after steaming, the greater the extent of retrogradation, and the less the degree of enzymatic digestibility. The decreased rate of enzyme digestibility correlated with amylopectin chain length distribution. Samples with short‐chain amylopectin exhibited a slow decrease in enzyme digestibility. It was determined that the structure and retrogradation properties of endosperm starch in Japanese rice cultivars affect the decreasing rate of enzyme digestibility of the steamed, milled rice grains.

海洋深層水が清酒酵母に与える影響

Deep Sea Water (DSW) can be pumped up from a depth of 200 meters at Muroto in Kochi. Since DSW is known to be unspoiled by pollution and contain various minerals, it is used for a variety of fermented foods. To investigate the effects of DSW on sake production, we carried out small-scale (1-2L) fermentation tests. As a consequence, the addition of DSW led to elevated levels of aroma components, including isoamyl acetate, ethyl caproate and ethyl caprylate. To elucidate further the mechanism underlying this phenomenon, we used cDNA microarray for studying yeast gene expression during fermentation. As a result, the addition of DSW substantially increased the transcript levels of several genes known to be involved in higher alcohol and fatty acid biosynthesis. It is therefore suggested that the rise in transcription of these genes caused the increased aroma production. Sensory evaluation revealed that the addition of DSW to fermentation resulted in a significantly superior flavor profile of final products compared with the addition of NaCl, a major component in seawater. Interestingly, the microarray analysis demonstrated that the addition of NaCl by itself induced stress during fermentation as indicated by mRNA levels in sake yeast. The addition of DSW appeared to ameliorate the negative effect induced by NaCl. Taken together, these results indicate that addition of DSW to the fermentation processes provides unique advantages for sake production.

Sequencing of Aspergillus nidulans and comparative analysis with A. fumigatus and A. oryzae

The aspergilli comprise a diverse group of filamentous fungi spanning over 200 million years of evolution. Here we report the genome sequence of the model organism Aspergillus nidulans, and a comparative study with Aspergillus fumigatus, a serious human pathogen, and Aspergillus oryzae, used in the production of sake, miso and soy sauce. Our analysis of genome structure provided a quantitative evaluation of forces driving long-term eukaryotic genome evolution. It also led to an experimentally validated model of mating-type locus evolution, suggesting the potential for sexual reproduction in A. fumigatus and A. oryzae. Our analysis of sequence conservation revealed over 5,000 non-coding regions actively conserved across all three species. Within these regions, we identified potential functional elements including a previously uncharacterized TPP riboswitch and motifs suggesting regulation in filamentous fungi by Puf family genes. We further obtained comparative and experimental evidence indicating widespread translational regulation by upstream open reading frames. These results enhance our understanding of these widely studied fungi as well as provide new insight into eukaryotic genome evolution and gene regulation.

Varietal Difference of Polishing Characteristics and Suitability for Sake Brewing in “Hattan-Type Varieties” of Rice Suitable for Brewing Original Hiroshima Sake

By successive crossing using Hattan-type varieties originating from “Hattanso” as a parent, “Hattan-type varieties” of rice suitable for brewing the original Hiroshima sake have been bred. The varieties were improved inheriting the flavor of Hattan-type sake, and Hattan-nishiki No.1 and No.2 were bred in 1984. However, neither variety was suitable for brewing high-grade sake such as Ginjoshu and Daiginjoshu, which require high-degree polishing of rice grains. Therefore, their parent cultivar, Hattan No. 35, is attracting attention for the production of high-grade sake. We have been trying to breed a new variety, which retains the sake-brewing suitability of Hattan-type varieties but can endure high-degree polishing. In this study, to establish a guideline for further breeding, we examined the polishing characteristics and suitability for sake brewing of six Hattan-type varieties derived from Hattanso. In the process of breeding of “Hattan-type varieties” of rice, grain size, white-core size and % of white-core grains increased resulting in an increased suitability for sake brewing, such as water absorptivity and digestibility, in Hattan No.35, Hattan-nishiki No.1 and Hattan-nishiki No.2. However, Hattan-nishiki No.1 and No.2 had many ellipsoidal-white-cores with large white tissue, which cause the grains to be easily broken during high-degree polishing. On the other hand, Hattan No.35, having grains with relatively many lined-white-cores with small white tissue, was superior for high-degree polishing. In the future, breeding of a new variety, which has the superior cultivation characteristics of Hattan-nishiki No.1 and No.2, but with improved white-core characteristics, is expected.

藩陽, 承徳, 北京での日本酒, 白酒などの生産と消費

[in Japanese]

焙煎発芽玄米の利用による高濃度アルコール清酒の製造

To develop high alcohol sake with the use of roasted germinated brown rice, we examined the process of sake production suitable for this purpose. Roasted germinated brown rice (RGB rice) was used as a replacement for tome-steamed rice at the test brewing. All the sake samples made with the addition of 5 to 10% RGB rice to the total rice were higher than the control (with no RGB rice) in contents of alcohol and the sake meter. Treatment with immersion and steaming of the RGB rice reduced the solubility of steamed rice compared with that receiving no treatment. However, the extracts of RGB rice powder by an acidic buffer (pH 4.0) inhibited the adsorption of α-amylase into the steamed rice. The sake made with the RGB rice was dry and light with high contents of alcohol.

清酒醸造の浸漬・蒸きょう工程における原料米α-グルコシダーゼの酵素反応

清酒醸造の浸漬・蒸きょう工程における原料米α-グルコシダーゼの酵素反応

Optimization of growth conditions of the postharvest biocontrol agent Candida sake CPA-1 in a lab-scale fermenter.

To maximize the growth (expressed as number of viable cells per millilitre) of the postharvest biocontrol agent Candida sake CPA(-1) at laboratory scale conditions. Growth conditons (aeration, agitation speed and inoculum size) were studied in batch conditions in a 5 l fermenter using molasses and urea as growth medium. Consumption of sugars and urea were analysed. Fed-batch studies were also carried out. Glucose and fructose were consumed during the exponential growth phase and were depleted after 18 h of growth. On the contrary, C. sake cells assimilated sucrose during the stationary phase. There was not growth improvement when fed-batch technology was used. Addition of an extra amount of glucose or molasses after 18 h of growth did not contribute to increase final population. Maximum growth (about 8 x 10(8 )CFU ml(-1)) was obtained at batch fermentation after 30 h growth at 400 rev min(-1), 150 l h(-1) of air and initial concentration of 106 CFU ml(-1). The results obtained in this study are an approach for further upscaling of C. sake production.

Molecular breeding of the Mureka-non-forming sake koji mold from Aspergillus oryzae by the disruption of the mreA gene

Mureka-non-forming sake koji molds were constructed from an Aspergillus oryzae industrial strain by the disruption of the mreA gene using a host-vector system with the ptrA gene as a dominant selectable marker. All of the mreA gene disruptants obtained retained the advantages of the host strain in terms of the brewing characteristics, while their isoamyl alcohol oxidase (IAAOD) activities were significantly lower than that of the host strain. Sake brewing was successfully carried out using the koji prepared with the disruptants, followed by storage of the resultant nonpasteurized sake (nama-shu). The isovaleraldehyde (i-Val) concentration in the sake brewed the host strain increased rapidly and reached the threshold values for mureka, 1.8 ppm and 2.6 ppm after storage at 20°C for 42 d and 63 d, respectively, while those of the disruptants were less than 0.5 ppm even after storage at 20°C or 30°C for 63 d. In the sensory evaluation of the sake sto red at 20°C or 30°C for 63 d, all members of the panel recognized the strong mureka flavor of the sake brewed with the host strain, while they did not detect this flavor in the sake brewed with the disruptants. Thus, we concluded that the mreA gene disruptants can be used for the production of sake in which mureka is not formed.

Uptake and removal of iron by immobilised persimmon tannin

Persimmon tannin was immobilised with formaldehyde (the aldehyde gel) and potassium peroxodisulfate (the persulfate gel). The resulting gels have high abilities to adsorb iron. The persulfate gel can adsorb iron from both iron(II) and iron(III) solutions with high efficiency. The adsorption of iron(III) by the persulfate gel was rapid and was affected by the solution pH, the concentration of iron in solution, and the mass of adsorbent. Adsorbed iron was easily desorbed with 0.1 mol dm−3 HCl solution. The gel can be repeatedly re-used for iron recovery using the adsorption–desorption cycle. The persulfate gel could remove trace amounts of iron in a sake-brewing water to facilitate the production of high quality sake. © 2000 Society of Chemical Industry

Breeding of a non-urea producing sake yeast with killer character using a kar1-1 mutant as a killer donor

Arginase-deficient (car1/car1) sake yeasts can brew sake without urea, a main precursor of ethyl carbamate, which is a suspected carcinogen in various fermented beverages. For the use of car1/car1 yeasts in sake production, contamination by wild-type (CAR1/CAR1) yeasts is a major problem. To protect sake mash against such contamination, killer character was introduced into the car1/car1 sake yeast HL163 by rare mating and protoplast fusion, using a kar1–1 haploid harboring killer dsRNA plasmids as a killer donor. All killer yeasts obtained showed no arginase activity and the same DNA content per cell as strain HL163, and produced sake with ordinary quality and very low levels of urea. We also demonstrated that one of these killer yeasts could effectively eliminate contaminant cells of a CAR1/CAR1 yeast from sake mash. Journal of Industrial Microbiology & Biotechnology (2000) 24, 203–209.

The Organization and Transformation of the Association of Sake Brewers in Saitama Prefecture during the Meiji Era

The aim of this paper is to examine how the association of sake brewers (syuzo-kumiai) was organized and expanded in modern Saitama. The trade association was an organization principally involved in the production and sale of sake. Research related to this field in Japan has focused on, first, how trade associations were established, although a cartel was banned by the Government at the beginning of modern era; second, why they increased in number; and, third, what were their main functions?A new entry into a cartel had been prohibited by the Shogunate until the end of the Edo era. The liberalization of business brought about a sudden increase in the number of sake brewers. Small newly-established sake brewers often attempted to evade liquor tax and to ensure bargain sales which caused confusion in the market. Meanwhile, large and medium-sized sake brewers organized the association of sake brewers (syuzo-kumiai) in 1875 to stabilize the market and restore order. However, the association of sake brewers did not perform well in Saitama Prefecture.The association grew into a nationwide organization to oppose a proposed tax increase in the 1880s. Although the campaign was initially supported by the opposition parties Jiyu-to and Kaisin-to, the former agreed to the necessity of a tax increase in 1896. As a result, the liquor tax was increased substantially in 1898, 1900, and 1902, respectively. The association of sake brewers (syuzo-kumiai) was obliged to halt its campaign since it had lost its raison d'etre.The price rise due to the tax increase reduced the demand for sake and resulted in strong competition within the industry. The sake brewers thus had to improve quality to overcome competition. Some members of the association of sake brewers established a brewing research laboratory, and held annual meetings to evaluate the quality of sake. The sake brewers who assiduously improved quality differed from those who had objected to the tax increase. The aims and achievements of each member of the association of sake brewers thus became quite diverse.

Uptake and removal of iron by immobilised persimmon tannin

Persimmon tannin was immobilised with formaldehyde (the aldehyde gel) and potassium peroxodisulfate (the persulfate gel). The resulting gels have high abilities to adsorb iron. The persulfate gel can adsorb iron from both iron(II) and iron(III) solutions with high efficiency. The adsorption of iron(III) by the persulfate gel was rapid and was affected by the solution pH, the concentration of iron in solution, and the mass of adsorbent. Adsorbed iron was easily desorbed with 0.1 mol dm−3 HCl solution. The gel can be repeatedly re-used for iron recovery using the adsorption–desorption cycle. The persulfate gel could remove trace amounts of iron in a sake-brewing water to facilitate the production of high quality sake. © 2000 Society of Chemical Industry

Isolation of sake yeast strains possessing various levels of succinate- and/or malate-producing abilities by gene disruption or mutation

Succinate and malate are the main taste components produced by yeast during sake (Japanese alcohol beverage) fermentation. Sake yeast strains possessing various organic acid productivities were isolated by gene disruption. Sake fermented using the aconitase gene ( ACO1) disruptant contained a two-fold higher concentration of malate and a two-fold lower concentration of succinate than that made using the wild-type strain K901. The fumarate reductase gene ( OSM1) disruptant produced sake containing a 1.5-fold higher concentration of succinate as compared to the wild-type, whereas the α-ketoglutarate dehydrogenase gene ( KGD1) and fumarase gene ( FUMI) disruptants gave lower succinate concentrations. The Δkgd1 disruptant exhibited lower succinate productivity in the earlier part of the sake fermentation, while the Δfum1 disruptant showed lower succinate productivity later in the fermentation, indicating that succinate is mainly produced by an oxidative pathway of the TCA cycle in the early phase of sake fermentation and by a reductive pathway in the later phases. Sake yeasts with low succinate productivity and/or high malate productivity was bred by isolating mutants unable to assimilate glycerol as a carbon source. Low malate-producing yeasts were also obtained from phenyl succinate-resistant mutants. The mutation of one of these mutant strains with low succinate productivity was found to occur in the KGD1 gene. These strains possessing various succinate- and/or malate-producing abilities are promising for the production of sake with distinctive tastes.

乾燥酵母協会701号および協会901号による清酒製造

We made two dry yeast starters of Saccharomyces cerevisiae strains K 701 and K 901.In order to make dry yeast these strains were grown aerobically in a cane molasses medium with a continuous supply of molasses, then the yeast cells were dehydrated in a dram dryer. Those dry yeast starters were used for SAKE brewing in four brewing companies on a production scale of approxinatly 3 ton of total rice weight.Every batch of fermented SAKE mash produced on excellent profile, and amino acidity did not increase even at the end of the fermentation period.This showed that these dry yeast strains aretolerant to a high concentration of ethanol.The SAKE produced had an excellent taste with a light and fruity fresh flavor.These results show that those dry yeast starters are useful for the practical production of SAKE.

酒類醸造およびパン製造のための有用酵母の育種

酒類醸造およびパン製造のための有用酵母の育種

A novel method for measurement of α-amylase activity in rice koji by capillary isoelectric focusing

A capillary isoelectric focusing (CIEF) method has been developed for the analysis of α-amylase activity in rice koji. The standards of α-amylase and rice koji extracts were successfully separated by CIEF in a large pH gradient. The α-amylase from A. oryzae was found to have pI of 3.7, as determined from the migration times of six standard proteins. A simple method has been developed for the measurement of α-amylase activity in rice koji, and the results were compared with those obtained by the official method. The coefficient of variation (C.V.) of the α-amylase assay was less than 0.5% in both intra-day and inter-day experiments. This CIEF method can be applied to the routine quality control of α-amylase activity in rice koji for the brewing process in sake production.