Increase In Germination Time Research Articles

Analyzing the effect of optimization conditions of germination on the antioxidant activity, total phenolics, and antinutritional factors of Chenopodium (Chenopodium album)

The purpose of present study was to optimize the effect of germination temperature and time on the total phenolic content, antioxidant activity, saponins, tannins and phytates in Chenopodium album grains using the response surface methodology (RSM). Statistical analysis revealed that independent variables (germination time and germination temperature) significantly (p < 0.05) affected all responses (AoxA, TPC, saponins, tannins and phytates). The results indicated that with increasing germination time and germination temperature, TPC and AoxA increased while the tannins, saponins and phytates decreased significantly (p < 0.05). The superlative combination of germination bioprocess variables for producing optimized flour with the higher TPC (5.64 g/100 g sample) and AoxA (18.45 g/100 g) while as lower saponins (0.67 g/100 g), tannins (1.29 mg/100 g), and phytate (0.46 mg/100 g sample), respectively with germination temperature (25.54 °C) and germination time of 49.59 h. Thus the results of this study showed that germination enhanced the TPC and AoxA while reducing the tannins, saponins and phytate of C. album flour and thus may help in improving its nutrient content. Therefore the optimized germinated C. album flour with high functional potential may find numerous applications in novel food formulations.

Purification and partial characterization of phytase from rice bean (Vigna umbellata Thunb.) germinated seeds

Rice bean (Vigna umbellata Thunb.) phytase activity increased during germination and reached maximum at 72 h. The phytate content in seeds decreased with increase in germination time. Phytase was purified 32 fold from 72-h germinated cotyledons with final specific activity 2.22 U/mg. Native PAGE revealed a single band. On SDS PAGE, it revealed two bands with molecular mass 66 and 44 kDa. The native molecular mass was 110 kDa on size exclusion chromatography. The A280/260 ratio was 1.88. When the enzyme was excited at 295 nm, the emission maximum was observed at 330 nm. The FTIR results suggest that Lys, Tyr, Phe, Trp, Ser, Gln and Asn residues on the enzyme’s surface. The enzyme was stored at 4 °C, showed 12 % residual activity on 35th day which was improved to 53.6 and 65.7 %, respectively in the presence of additives ascorbic acid and acetaminophen. The optimum pH and temperature of enzyme were 4.0 and 40 °C, respectively. The energy of activation was 32.2 kJ/mol. The values of Km and Vmax were 0.197 mM and 2.35 μmol/min/mg protein, respectively with sodium phytate as substrate. Phytase showed broad substrate specificity. The kcat/Km ratio was the highest for sodium phytate.

발아 전후 보리 당단백질 추출물의 영양성분 및 면역 활성 변화

In the present study, we investigated changes in the nutritional components of pre- and post-germinated barley seeds and also investigated their immune-enhancing effects such as production of nitric oxide (NO), interleukin (IL)-6, and tumor necrosis factor (TNF)-α on the RAW 264.7 macrophage cell line. Protein and total sugar contents increased slightly with increase in germination time. The major neutral sugars of germinated balrey seeds were arabinose, glucose, and xylose. Glucose content decreased during germination, whereas arabinose and xylose contents increased during germination. Amino acid contents of barley germinated for 24 and 48 hours increased 1.03-fold and 1.24-fold, respectively, compared to that of pre-germinated barley. Moreover, RAW 264.7 macrophages stimulated with barley germinated for 24 and 48 hours showed higher production of NO, IL-6, and TNF-α compared to that observed in pre-germinated barley. The results of the present study indicate that germinated barley may have immune-enhancing effects derived from its ability to activate RAW 264.7 macrophages, which play a major role in innate immunity.

발아시간에 따른 팥의 항산화성분 및 항산화활성의 변화

본 연구에서는 나물용 팥으로 새롭게 개발된 팥 신품종 연두채의 나물로의 이용 가능성을 평가하고 그 추출물에 대한 항산화성분 및 항산화활성을 측정하였다. 발아시간에 따른 싹 및 뿌리의 길이와 나물 수율을 고려해 볼 때 연두채는 120~144시간 발아시키는 것이 바람직할 것으로 생각된다. 발아시간에 따라 수분은 유의적으로 증가하였고 단백질은 유의적으로 약간 증가하였으나 큰 차이를 보이지 않았다. 연두채의 칼슘, 마그네슘 함량은 유의적으로 증가하는 경향을 보였고 칼륨, 나트륨, 알루미늄 함량은 유의적인 차이를 보이지 않았다. 발아시간에 따른 에탄올 추출물의 총 폴리페놀, 플라보노이드 및 탄닌 등 항산화성분의 함량은 발아시간에 따라 유의적인 차이를 보였다. 충주팥, 연두채, 다현녹두원료곡의 총 폴리페놀 함량은 각각 1.96, 2.68 및 2.02 mg/g이었고 충주팥과 연두채는 144시간(각각 3.33 및 3.47 mg/g)에 높은 함량을 나타내었다. 원료곡의 총 플라보노이드 함량은 발아시간이 증가함에 따라 충주팥과 연두채가 대체적으로 감소하는 경향을 보였다. 원료곡의 총 탄닌 함량은 발아시간이 증가함에 따라 대체적으로 증가하는 경향을 보였고 충주팥은 120시간(0.96 mg/g), 연두채는 144시간(0.85 mg/g), 다현녹두는 120시간(1.62 mg/g)에 높은 함량을 보였다. 원료곡의 DPPH radical 소거활성은 발아시간이 증가함에 따라 충주팥과 연두채는 대체적으로 감소하고 다현녹두는 증가하였다. ABTS radical 소거활성은 각각 4.57, 6.51 및 2.82 mg TE/g이었고 충주팥과 연두채는 발아 초반에 감소하다가 각각 120 및 72시간 이후 증가하였다. 이상의 결과 연두채 싹나물은 다현녹두 나물에 비해 항산화성분 및 활성은 약간 낮으나 수율이 높고 탄닌 등 쓴맛 성분이 낮아 싹나물로의 이용성이 기대된다.

Effect of Germination on Functional Properties and Degree of Starch Gelatinization of Sorghum Flour

&lt;p&gt;Sorghum grains were germinated for 24, 48 and 72 hours with a view to determining the effect of germination on some functional properties and degree of starch gelatinization of the flour. Flour from non-germinated grains served as control. In order to measure the effect of germination on degree of starch gelatinization, the flours were processed into cookies. Germination of sorghum grains for 48 hours and above significantly (p&amp;lt;0.05) decreased both loose and packed bulk densities from 0.59 g/ml and 0.77 g/ml to 0.56 g/ml and 0.70 g/ml respectively. The water absorption capacity of the sample germinated for 72 hours was 1.38 g/g which was significantly (p&amp;lt;0.05) higher than the other samples. The oil absorption capacity of the samples germinated for 48 and 72 hours (1.16 and 1.18 g/g respectively) were significantly (p&amp;lt;0.05) higher than those of the control sample and 24 hour germination (1.03 and 1.04g/g respectively). Germination also significantly (p&amp;lt;0.05) increased the swelling power (22-23.2 ml/g), foaming capacity (14-16.2%) and emulsion capacity (58.6-65.5%). The degree of starch gelatinization increased with increasing germination time but decreased with increasing temperature. Generally, germination had a beneficial effect on the functional properties measured. Flour obtained from sorghum grains germinated for 72 hours had the best results.&lt;/p&gt;

Influence of Elicitation and Germination Conditions on Biological Activity of Wheat Sprouts

The influence of elicitation, time, and temperature of germination on the biological activity of wheat sprouts was studied. The total phenolics content (TPC), antioxidant activity (AA), and microbiological purity of seeds of three wheat cultivars elicited withSaccharomyces cerevisiae(Y) andSalix daphnoidesbark extracts (W) and their mixture (1 : 1) germinated in different conditions were studied. The increase of germination time caused an increase the TPC and AA for all tested wheat cultivars. Germination temperature also influenced these parameters. Generally the highest values in TPC were obtained for sprouts germinated at 20°C. This tendency was found for all tested wheat cultivars. The highest values of TPC and AA were obtained after elicitation byS. daphnoidesbark extract. The results of microbiological examination showed that generally for seeds germinated at lower temperature (15°C and 20°C) and during 2 and 4 days the molds were not detected and the levels of aerobic bacteria did not exceed the usually accepted limits. Taking into account both biological activity and microbiological purity of samples the best quality sprouts were in the most cases obtained for wheat germinated during four days at 20°C and elicited by W.

Changes of Tocopherols, Tocotrienols, γ-Oryzanol, and γ-Aminobutyric Acid Levels in the Germinated Brown Rice of Pigmented and Nonpigmented Cultivars

This study examined the changes of tocopherols (Toc), tocotrienols (T3), γ-oryzanol (GO), and γ-aminobutyric acid (GABA) contents in germinated brown rice (GBR) of pigmented and nonpigmented cultivars under different germination conditions. Results showed that the Toc and T3 contents in GBR were significantly different between treatments in both rice cultivars. The pigmented GBR possessed higher total vitamin E, total Toc, total T3, and GO contents than the nonpigmented GBR; however, its level of GABA was lower. The order of the three highest vitamin E homologues in pigmented and nonpigmented GBR was γ-T3 > γ-Toc > α-Toc and α-Toc > γ-T3 > α-T3, respectively; β-Toc, β-T3, δ-Toc, and δ-T3 were present in only small amounts (≤1.0 mg/kg) in GBR of both cultivars. Although both cultivars showed an increase in GABA contents with increasing germination time, the GABA content in nonpigmented GBR was higher.

Major nutrient compositions and functional properties of sorghum flour at 0–3 days of grain germination

Germination is of importance to improving nutritional attributes of cereal grains for human consumption. The effect of germination time on major nutrient compositions and functional properties of sorghum flour was investigated in this study. Grains of Butanua, a new Sudanese sorghum cultivar, were germinated for 0, 1, 2, and 3 days to analyze their chemical and functional properties. The contents of starch, protein, oil, foaming stability, bulk density, and least gelation concentration of sorghum flour decreased, whereas oil absorption capacity, foaming capacity, and emulsion capacity and stability enhanced with an increase in germination time. Improved functional properties of sorghum flour by germination of the grains not only make it useful and suitable for various food processing formulations, but also improve the food product quality. This new finding will beneficially help develop innovative technologies, design new types of functional foods, and promote both sorghum production and relevant food processing industry in the future.

Effects of Bonny light crude oil contamination on the germination, shoot growth and rhizobacterial flora of Vigna unguiculata and Arachis hypogea grown in sandy loam soil

This study was carried out to evaluate the effects of different levels of Bonny light crude oil contamination on the germination, shoot growth and rhizobacterial flora ofVigna unguiculata (cowpea) and Arachis hypogea (groundnut) grown in potted sandy loam soil samples in Nsukka, Nigeria. Crude oil at a level of 2.5% increased germination time in cowpea by 24 h but at higher doses (5 to 20%) germination of cowpea seeds was totally inhibited. On the contrary, germination of groundnut seeds occurred at all levels of crude oil pollution although at different rates. The germination time was 7 days in soils with 0.5 to 2.5% oil concentration and an average of 9 days in those with 5 to 20% levels of the pollutant. Even though groundnut germinated and grow in all concentrations of crude oil tested, significant (p<0.05) shoot growth retardation still occurred in both legumes consequent on crude oil toxicity. Rhizobacterial population also diminished with increase in crude oil concentration. The study reveals both the vulnerability of cowpea and the resistance of groundnut to crude oil, marking groundnut out as a promising phytoremediation candidate. Key words: Crude oil, pollution, rhizobacteria, legume, phytoremediation, rhizosphere.

Storage Proteins and Trypsin Inhibitors of an Underutilized Legume, &amp;lt;i&amp;gt;Mucuna&amp;lt;/i&amp;gt;: Variability and Their Stability during Germination

The proteins and trypsin inhibitors were isolated from the seeds of different varieties/accessions of an underutilized legume, Mucuna. The crude protein content of all the germplasms of Mucuna is varied from 15% - 26%, showed little variation and contain higher crude protein when compared with other Mucuna species reported earlier and the pulse crops commonly consumed in India. The seeds of all the varieties of Mucuna exhibited trypsin inhibitor activity. The trypsin inhibitor activity varied from 11 - 14 TIA/mg of protein. Not much variation was observed in trypsin inhibitory activities in soaked seeds compared to dry seeds. Germination of Mucuna pruriens has been carried out and the change in the protein content and trypsin inhibitors were monitored. The protein content of the endosperm increased up to 72 hrs of germination and then decreased. The trypsin inhibitory activity decreased with increase in germination time. The trypsin inhibitor activity was decreased from 14.81 TIA/mg to 2.62 TIA/mg (82% reduction in the trypsin inhibitor activity) after 144 hrs germination.

Mineral bioavailability in three locally consumed pulses processed using popular methods: interpreted using molar ratios with phytic acid

Pulses, an essential component of the Sri Lankan diet, have high levels of phytic acid (PA) which chelate essential mineral nutrients limiting their bioavailability. Three locally grown pulses; mung beans, black gram and soya bean were investigated to determine the effects of popular processing methods on their Zn, Fe and Ca bioavailability as affected mainly by phytic acid (PA). Processed mung beans (Vigna radiata var. MI 6) and black gram (Vigna mungo var. MI 1) had very high levels of PA. In mung beans, with increasing germination time, a significant decrease (P=0.05) in PA from 2517 to 1998 mg per 100g (dry weight basis) was observed but PA:Zn molar ratio did not improve. Natural fermentation of black gram, did not affect PA level effectively, but levels decreased when inoculated with a lactic acid bacterium or with Pantoea agglomerans. In soya bean (Glycine max var. Pb), significant (P=0.05) reduction in PA was observed in tempeh (487 mg/100 g dry weight). Overall, in soya products, the Zn bioavailability has increased from low to moderate levels in both tofu and tempeh. The molar ratio of PA:Fe was lower in all soya products. Although, PA:Zn molar ratio has decreased in germinated mung beans, the Ca x PA:Zn ratio exceeded 0.5 indicating that Ca influences Zn bioavailability DOI: http://dx.doi.org/10.4038/cjsbs.v41i1.4534 Ceylon Journal of Science (Bio. Sci.) 41 (1): 19-26, 2012

Effect of germination on antinutritional factors, total, and extractable minerals of high and low phytate corn (Zea mays L.) genotypes

Two corn genotypes, Var-113 (high phytate) and TL-98B-6225-9×TL617 (low phytate) were germinated for 6days. The germinated seeds were dried and milled. Phytic acid, polyphenols, and hydrochloric acid (HCl) extractable minerals were determined at intervals of two days of germination. Phytic acid decreased significantly (P⩽0.05) with a concomitant increase in HCl extractable minerals, while polyphenol contents increased significantly (P⩽0.05) with an increase in germination time. However, the major mineral content was initially decreased and then increased while that of trace minerals was increased with germination time. After 6days of germination, the high phytate sample (Var-113) exhibited higher extractable calcium compared to the other genotype, whereas the low phytate sample (TL-98B-6225-9×TL617) showed higher extractable phosphorus. Iron extractability was higher for the low phytate samples, while that of magnesium was higher for the high phytate sample. High correlation between lower phytate and higher extractable minerals was noted as a function of longer germination time. Nevertheless, no correlation was observed between polyphenol content and mineral extractability level.

Optimization of germination time and heat treatments for enhanced availability of minerals from leguminous sprouts.

Germinated legumes are highly nutritious food especially for their enhanced iron bioavailability primarily because of reduction of phytates and increase in ascorbic acid with an advancement of germination period. Length of germination time followed by different heat treatments affect the nutritive value of leguminous sprouts. To optimize germination time and heat treatments for enhanced availability of iron from leguminous sprouts, three legumes namely, mungbean, chickpea and cowpea were germinated for three time periods followed by cooking of sprouts by two cooking methods ie. pressure cooking and microwaving. Optimized germination time for mungbean was 12, 16 and 20h; 36, 48 and 60h for chickpea and 16, 20 and 24h for cowpea. Germination process increased ascorbic acid significantly in all the three legumes, the values being 8.24 to 8.87mg/100g in mungbean, 9.34 to 9.85mg/100g in chickpea and 9.12 to 9.68mg/100g in cowpea. Soaking and germination significantly reduced the phytin phosphorus in all the three legumes, the percent reduction being 5.3 to 16.1% during soaking and 25.7 to 46.4% during germination. The reduction in phytin phosphorus after pressure cooking was 9.6% in mungbean, 18.4% in chickpea and 6.1% in cowpea. The corresponding values during microwaving were 8.4, 19.7 and 4.5%. Mineral bioavailability as predicted by phytate:iron enhanced significantly with an increase in germination time. Further reduction i.e. 0.9 to 16.3% was observed in three legumes after the two heat treatments. The study concluded that the longer germination periods ie. 20h for mungbean, 60h for chickpea and 24h for cowpea followed by pressure cooking for optimized time were suitable in terms of better iron availability.

Impact of Low Hydration of Barley Grain on β-Glucan Degradation and Lipid Transfer Protein (LTP1) Modifications During the Malting Process

One of the objectives of the malting industry is to reduce the energy cost during kilning without major effect on malt quality. In this study, the impact of a low hydration steeping process on lipid transfer protein (LTP1) modifications and β-glucan breakdown was evaluated in low (LH) and high (HH) hydrated malts. LTP1 modifications analyzed by MS/MS revealed acylation, glycation, and disulfide bond breakage in both LH and HH malts. LTP1 free amine content measurement and fluorescence of Maillard protein adducts revealed no significant difference between LH and HH malts. Immunolabeling of LTP1 during malting highlighted the diffusion of the protein from the aleurone layer to the endosperm at the end of steeping in both LH and HH malts. By contrast, a significant higher amount of β-glucans was measured in LH malts after five days of germination, whereas no significant difference between LH and HH malts was revealed through immunostaining of β-glucans or evaluation of the endosperm integrity after seven days of germination. The possibility to reduce the effects of a low hydration steeping process on β-glucan hydrolysis by increasing germination time was discussed.

Selenium accumulation in protein fractions during germination of Se-enriched brown rice and molecular weights distribution of Se-containing proteins

This study investigated the accumulation of selenium (Se) in protein fractions of albumin, globulin, prolamin and glutelin extracted from Se-enriched brown rice and the molecular weight distribution of Se-containing proteins. Results showed that the amount of total Se (T-Se) and protein-bound Se (PB-Se) in brown rice was significantly ( P < 0.05) increased after germination with 10–60 μmol/l sodium selenite. Except prolamin, the amount of all the other three protein fractions decreased significantly ( P < 0.05) with the increase of germination time. Low Se concentrations had promoting effects on degradation of albumin and globulin, while no significant effects were observed on prolamin and glutelin. The accumulation of T-Se and PB-Se were in the order of albumin > glutelin > globulin > prolamin. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS–PAGE) analysis showed that Se was distributed in all the proteins of which molecular weights varied from 13.6 to 121.4 kDa; however, 84.34% of Se was observed in the proteins whose molecular weights less than 36.3 kDa.

The effects of malting conditions on the diastatic power of three malted Nigerian sorghum cultivars

The effects of malting conditions on the diastatic power of three malted sorghum cultivars (SK 5912, ICSV 400 and KSV 8) were investigated. The three Nigerian sorghum cultivars were steeped in 0.01M NaOH, 0.01M Ca(OH)2 and distilled water, respectively, under air-rest and continuous steeping regimes. SK 5912 steeped in Ca(OH)2 gave higher hot water and cold water extract values than when steeped in NaOH and distilled water. KSV 8 when steeped in distilled water gave peak values for cold water extract during air-rest and continuous steeping regimes. For hot water extract, KSV 8 had peak values when steeped in NaOH whereas SK 5912 gave the least values when steeped in distilled water. Cold water soluble-carbohydrate gave peak values for ICSV 400 when steeped in Ca(OH)2 and the least values were recorded in NaOH steeped grains. The diastatic power of the three sorghum cultivars improved with increase in germination time except for ICSV 400-steeped in Ca(OH)2 and KSV 8 steeped in distilled water which showed decrease in activity on the 4th day in continuous steeping regime. The results show that steeping in dilute alkali solutions gave higher values than distilled water while air-rested steeping regime gave better values than continuous steeping regime. The results from this study suggest that increase in germination time enhanced the diastatic power of the malted grain. Keywords: Sorghum varieties, Malting, Diastatic power, Cold water extract, Hot water extract

Influence of grain germination on functional properties of sorghum flour

A Sudanese sorghum cultivar (Fetarita) was germinated for five days and protease and amylase activities were measured every 24 h. Results showed that flour obtained from the 3rd germinated sorghum grain had high protease and amylase activities. The functional properties of flours derived from the germinated sorghum seeds were studied and ungerminated seeds were used as a control. Germinated samples had a higher protein solubility compared with the control, and the highest solubility occurred at pH 6. Germination also increased the protein solubility index of sorghum flour. Germinated sorghum flour had a least gelation concentration of 8% compared with 18% for the control. The bulk densities of germinated flours were lower compared to the ungerminated one. Water and oil capacities were increased by germination from 131.34% and 90.56% to 141.64% and 108%, respectively after three days of germination. The emulsifying activities and stabilities of the germinated samples increased significantly. In addition, germination improved the foamability of sorghum flour from unfoam flour to a flour with foam after three days of germination; and the foaming capacity and stability increased significantly with increasing germination time. Thus, the study indicated that germination improved the functional properties of sorghum and it would be possible to design new foods using germinated sorghum.

Effect of time and temperature on bioactive compounds in germinated Brazilian soybean cultivar BRS 258

Abstract The objective was to determine the effect of time and temperature on the concentration of bioactive compounds during germination of Brazilian soybean cultivar BRS 258. The concentration of bioactive compounds was optimized using Response Surface Methodology, with a 22 central composite rotational design and germination time and temperature as independent variables. Germination was carried out in a germination chamber, using paper in trays containing 500 g of seeds. Germination temperature and time modified the concentrations of bioactive compounds within the ranges studied. An increase in germination time at 25 °C decreased the concentration of Bowman–Birk inhibitor, lectin and lipoxygenase. After 63 h of germination, a temperature increase from 20 °C to 30 °C resulted in a decrease of lipoxygenase activity by 22.5%. Optimal increases in the concentrations of isoflavone aglycones (daidzein and genistein) and saponin glycosides were observed with a 63 h germination time at 30 °C. Both germination time and temperature had an influence on the composition and concentration of bioactive compounds in germinated soybean flour. Equations determined can be used to predict concentrations of bioactive compounds in germinated soybeans in relation to climate changes and thus optimize their potential use in human health and nutrition.

Effect of pretreatments on seed viability during fruit development of two varieties of &lt;i&gt;Irvingia gabonensis&lt;/i&gt;

Studies to identify the stage at which developing fruits of Irvingia gabonensis (var. excelsa and var. gabonensis), picked from standing trees and/or forest floors, attain maximum viability and germinability were conducted in two harvesting seasons in 2000 and 2001. Some pretreatment methods were used as a means of stimulating expression ofmaximum germinability as source of seeds for seedling production. The moisture contents of whole seeds decreased steadily and significantly (P = 0.05) throughout the developmental stages of both varieties and in both harvests. Seeds harvested from standing trees and forest floors, pretreated by steeping in water for 24 h at 26 oC and sown in sawdustand petri dishes on moist filter paper, gave 80% germination in variety gabonensis, 20 weeks after fruiting. Germination potentials of seeds in both harvests were enhanced significantly (P = 0.05) with seeds pretreated by steeping in water for 24 h at 26 oC, and sown in sawdust and petri dishes on moist filter paper by as much as 60% and 80% for var. excelsa and var. gabonensis, respectively, in the developmental stages, 14–20 weeks after fruiting with significant (P = 0.05) decrease in germination time to 21 and 15 days, respectively. There was also significant (P =0.05) relationship between viability and germination in both harvests and in the two varieties throughout the developmental stages. Potassium nitrate (KNO3) significantly (P = 0.05) enhanced germination potentials of the seeds over the control when soaked in 1.5 g/l concentration in both varieties and in the two harvests (18–20 WAF) by 80%. Mannitol significantly (P = 0.05) reduced germination potential (40%) of the seeds with increased germination time in both harvests and in the two varieties throughout the developmental stages. There was a high significant (P = 0.05) increase in germination potential (60%) of seeds treated with polyethylene glycol (PEG) when soaked in concentration of 10 g/l for 48 and 72 h in both varieties and in the two harvests (18–20 WAF). The study shows that the fruits of I. gabonensis assume ripening characteristics (yellowing) and that the viable seeds can be obtained from standing treesand/or picked from forest floors from the 12th to 20th week after fruiting.

Effects of steeping and anaerobic treatment on GABA (γ-aminobutyric acid) content in germinated waxy hull-less barley

Effects of steeping conditions (time, temperature and soaking solution) and anaerobic storage on the gamma-aminobutyric acid (GABA) content in waxy hull-less barley grains during germination was examined. The barley kernel was steeped for 16 h at different temperatures (5, 15 or 35 °C) either in water or in a buffer solution (pH 6.0, 50 mmol/L sodium acetate) and then germinated at 15 °C for 72 h. To reach the optimum water content (36–44 g/100 g) for germination, a longer steeping period was required when steeping temperature was lower (16 h at 5 °C vs. 8 h at 15 °C). At 35 °C for steeping, however, the water content in the grains increased excessively, and thus germination percentage became much less than those at 5 and 15 °C. The GABA content increased with increasing germination time and was higher in the buffer solution than water. These findings indicate that the glutamate decarboxylase (GAD), which is the rate-limiting enzyme for GABA synthesis, is more activated by extending germination at controlled pH (6.0). An anaerobic storage with nitrogen in the dark for the germinated barley grains substantially raised the GABA content: 14.3 mg/100 g after the treatment for 12 h, which was four times higher than that of control sample (3.7 mg/100 g). Overall results suggest that the steeping prior to germination greatly affects the GABA production during the germination of barley, and the anoxia storage with nitrogen after the germination increases the GABA content.