Fermented Fish Sauce Research Articles

Role of jeotgal, a Korean traditional fermented fish sauce, in microbial dynamics and metabolite profiles during kimchi fermentation

We investigated the effects of jeotgal (fermented fish sauce) on kimchi fermentation, with or without saeu-jeot and myeolchi-jeot. Bacterial community analysis showed that Leuconostoc, Weissella, Lactobacillus, and Tetragenococcus were the dominant genera; however, their succession depended on the presence of jeotgal. Leuconostoc gasicomitatum was the dominant species in kimchi without jeotgal, whereas Weissella koreensis and Lactobacillus sakei were the dominant species in kimchi with myeolchi-jeot and saeu-jeot, respectively. Metabolite analysis, using 1H NMR, showed that the amounts of amino acids and gamma-aminobutyric acid (GABA) were higher in kimchi with jeotgal. Increases in acetate, lactate, and mannitol contents depended on fructose consumption and were more rapid in kimchi with jeotgal. Moreover, the consumption of various amino acids affected the increase in kimchi LAB. Thus, the role of jeotgal in kimchi fermentation was related to enhancement of taste, the amino acid source, and the increases in levels of functional metabolites.

ISOLATION OF HALOPHILIC LACTIC BACTERIA TETRAGENOCOCCUS HALOPHILUS FROM VIETNAMESE FISH SAUCE

Halophilic lactic bacteria Tetragenococcus halophilus were isolated from Thailand fish sauce. They were suggested to play an important role in flavor and aroma development and used as starter culture during fish sauce fermentation.In this study the isolation of Tetragenococcus halophilus from two different factories of fish sauce production in Vietnam was conducted on MRS agar media supplemented with 0.5 % CaCO3 and 5 % NaCl under anaerobic condition. The Tetragenococcus halophilus was present in all investigated fish sauce samples from 2 months to 8 months. Six isolates CH2-4, CCH8-3, CH6-1, CH6-2, CH8-1 and V5-1 were selected based on various ability to grow in fish broth supplemented with 25 % NaCl and degrade fish protein to oligopeptide. The 16S rDNA sequence analysis of 6 isolates showed that they were closely related to Tetragenococcus halophilus ATCC 33315 and Tetragenococcus halophilus MCR10-7-8 and MRC 5-5-2 from study of Thailand scientist with more than 99 % sequence homology, suggesting their potential using as starter culture in fish sauce fermentation.

Effect of Bacterial Community and Free Amino Acids on the Content of Biogenic Amines During Fermentation of Yu-lu, a Chinese Fermented Fish Sauce

ABSTRACTThe main objective of this work was to investigate the effects of bacterial community and free amino acids on the content of biogenic amines in Yu-lu during fermentation. Four major biogenic amines (histamine, tyramine, cadaverine, and putrescine) were identified, and the content of tyramine was positively correlated with histamine (R value = 0.9113). Aspartic acid, glutamic acid, histidine, leucine, and lysine were the dominant free amino acids. 16S rRNA sequencing showed that the composition of the bacterial community changed significantly during fermentation of Yu-lu. Principal component analysis revealed the crucial links between microbial community and biogenic amines. For example, Halanaerobium was probably associated with the formation of putrescine, while Halomonas might be associated with the degradation of biogenic amines at the end of fermentation of Yu-lu. This study provided a detailed evaluation of the Yu-lu fermentation process, enabling development of better strategies for biogenic amine control in fish sauce.

Identification of sensory characteristics that drive consumer preferences of commercially mass‐produced doenjang in Korea

AbstractThe objective of this study was to identify the drivers of liking for commercially mass‐produced doenjang (fermented soybean paste) among Korean consumers, by correlating the results from descriptive analysis and consumer acceptance testing. Ten doenjang samples were tested, seven of which were manufactured through a modified method and three via applying traditional method for mass production. Significant differences were found in all sensory attributes among the samples (p < .05). The principal component analysis of the descriptive analysis revealed clear discrimination among doenjang samples, according to their sensory characteristics: one group favored mild flavors (high in “sweet” and “umami” tastes and “cooked soybean flavor,” while the other preferred strong fermentation‐related flavors, such as “musty,” “biting,” “soy sauce,” and “fermented fish sauce.” Overall, consumer preference was directed toward doenjang that is high in sweet and umami tastes, meaning that customers favored doenjang produced by the modified method more than traditional‐style doenjang.Practical applicationsThe doenjang market is currently greatly expanding, as consumer interests in health and traditional foods are increasing. Therefore, it is more important than ever to identify the sensory characteristics driving consumer preference, in order to meet consumer expectations. The present study identified consumers who preferred traditional doenjang characteristics as well as sweet and umami characteristics. This suggests the direction in which to proceed in optimizing doenjang flavors, in order to ensure that they match Korean consumer preferences.

Flavour of fermented fish, insect, game, and pea sauces: Garum revisited

Fermentation of dairy products, legumes, fish, shellfish, molluscs, and meat are considered to lead to some of the most flavourful products in cuisines across the world. The deliciousness of these fermented products is to a large extent due to compounds like free amino acids and free nucleotides, formed during fermentation, which impart umami taste, often in a synergistic fashion. We have prepared fermented fish sauces based on mackerel, using procedures as in the ancient Roman cuisine, and used similar techniques to produce experimental fermented sauces from insects (moths and grasshoppers), game (pheasant), and pulses (peas). In some cases, the fermentation has been facilitated by fungal inoculation based on a Japanese koji mother. We have performed chemical analysis of these experimental fermentation products, together with a comparative analysis of a series of commercial fish sauces, with particular focus on free amino acids and free nucleotides in order to assess the umami potential of the various products. Whereas all of the 21 different investigated sauces are found to have high amounts of glutamate and aspartate, no significant amounts of free nucleotides are found in any of the samples. The investigated sauces were characterized by quantitative sensory evaluation. Although high in glutamate, umami synergy is not expected to play any significant role for the flavour of these fish, insect, game, and pea sauces. The sensory analysis shows a fairly good prediction of sensory properties from the chemical characterization of the sauces. However, the relationship between glutamate/aspartate concentration and intensity of umami taste is not simple. It demonstrates that in the complex solutions that constitute these sauces, there may be other perceptions that interfere with the main umami-tasting compounds.

Understanding the sensory characteristics and drivers of liking for gochujang (Korean fermented chili pepper paste).

The present study investigated the sensory characteristics of 31 types of gochujang, a Korean fermented chili paste, produced in various regions of Korea. Generic descriptive analysis was conducted to analyze the sensory properties of the samples. Among these, four samples were selected for consumer taste test based on their distinct sensory profiles. The drivers of liking for gochujang were identified when the samples were applied to four different food systems. Consumers (age range: 20's-60's) rated the acceptance of the samples. ANOVA and multivariate analyses were conducted on the data. The types of carbohydrate sources or usage of additional seasoning showed stronger influence on the sensory characteristics of gochujang than the production location. Samples with strong fermented soy and fish sauce characteristics were fairly unacceptable to the consumers. The positive drivers of gochujang were appropriate levels of spiciness and sweet odors/flavors, whereas the negative drivers were off-note and fermentation-related flavors.

Isolation and identification of halophilic bacteria producing halotolerant protease

Halotolerant proteases are known as one of the important groups of enzymes that have been used widely in various industries. However, high production cost of proteases in term of energy used for sterilization and high risk of microbial contamination during fermentation become the obstacles during upstream processing. Moreover, proteases that capable to withstand with harsh conditions of salinity, temperature and pH are required in industry. Therefore, this study presents newly isolated halophilic bacteria producing halotolerant proteases. Halophilic bacteria were isolated from fermented fish sauce. Screening of the bacteria producing halotolerant protease enzymes was carried out by using skim milk salt agar containing 10% NaCl at pH 7 and incubated under aerobic condition at 37oC for 2 days. The selected isolates were identified based on their morphology followed by 16S rDNA gene sequence analysis. 40 colonies of halophiles bacteria were isolated, however, only 20 of them showing proteolytic activity. All of 20 isolates are motile and gram positive bacteria. From 20 isolates, only 6 were chosen for further analysis. B7 showed the highest proteolytic activity compared with others. Results of 16S rDNA gene sequence analysis showed 98% homology to Bacillus amyloliquefaciens subsp. plantarum strain FZB42. Therefore, B7 is identified as Bacillus amyloliquefaciens strain B7.

Improvement of fish sauce quality by combined inoculation of Tetragenococcus halophilus MS33 and Virgibacillus sp. SK37

Biological and chemical dynamics were investigated during fermentation of fish sauce inoculated single and combined starter cultures, Virgibacillus sp. SK37 and Tetragenococcus halophilus MS33. At 120–150 days of fermentation, cell numbers of Virgibacillus sp. SK37 and T. halophilus MS33 were lower as detected by qPCR in fish sauce inoculated with a single culture compared with the sample inoculated with combined cultures. In addition, qPCR revealed that the sequential inoculation of Virgibacillus sp. SK37 followed by the addition of T. halophilus MS33 after 1 month (SK37_1M + MS33) showed bacterial counts that were approximately 0.5–1 LogCFU/mL higher than the single starter culture treatments. α-Amino contents of fish sauce added simultaneous co-cultures (MS33 + SK37) and SK37_1M + MS33 were 1160.41 and 1185.71 mM, respectively. SK37_1M + MS33 showed a lower amount of histamine than the control (P < 0.05). Fish sauce samples inoculated with either sequential cultures or a single culture of Virgibacillus sp. SK37 or T. halophilus MS33 showed higher amounts of total glutamic acid and greater total amounts of amino acids than the control (P < 0.05). The amounts of key desirable volatiles, namely 2-methylpropanal, 2-methylbutanal, and 3-methylbutanal, were higher in SK37_1M + MS33 than in the control (without starter culture, P < 0.05). The addition of both starter cultures in sequential order showed the potential to improve quality in terms of volatile compounds, glutamic acid content and overall acceptability.

Statistical optimization for the improved production of an extracellular alkaline nuclease by halotolerant Allobacillus halotolerans MSP69: Scale-up approach and its potential as flavor enhancer of fish sauce

Abstract An efficient extracellular alkaline nuclease producing halotolerant bacterium was isolated from fermented shrimp paste ( kapi ) and identified as Allobacillus halotolerans based on the phenotypic analysis and 16S rDNA sequence homology. The temperature and pH ranges for growth were 20–45 °C (optimum at 37 °C) and pH 6.0–9.0 (optimum at pH 8.0). A statistical optimization method was applied to enhance the alkaline nuclease production using a 20-run, 3-factor with 5-level of the central composite design (CCD) generated by design expert software. The optimum conditions for the simultaneous production of deoxyribonuclease (DNase) and ribonuclease (RNase) in a single media were determined as 17.5 g/L of yeast extract, 1.59% (w/v) NaCl and an initial pH 8.0. The maximum productions of RNase (59.36 U/mL) and DNase (141.21 U/mL) were in agreement with the prediction values and the model was proven to be adequate. The optimized conditions from the shake-flask experiments were validated in a 7 L lab scale fermenter, and the RNase and DNase production increased to 3.95-fold and 6.77-fold, respectively. Furthermore, the alkaline nuclease preliminary employed as flavor enhancer in the fish sauce fermentation process increased the aspartic acid (776.03 mg/100 mL), glutamic acid (1613.78 mg/100 mL), 5′-GMP (36.89 mg/100 mL) and 5′-AMP (21.49 mg/100 mL) in fish sauce ( P

Purification and characterisation of a salt-stable protease from the halophilic archaeon Halogranum rubrum.

Because proteases play an important role in the fermentation of fish sauce, the purification and characterisation of an extracellular protease from the halophilic archaeon Halogranum rubrum was investigated. The molecular mass of the protease was estimated to be approximately 47 kDa based on sodium dodecyl sulfate-polyacrylamide gel electropheresis (SDS-PAGE) and native-PAGE analysis. The optimum conditions for catalytic activity were pH 8.0 and 50°C. The protease showed alkaline stability (pH 7.0-10.0). The protease also exhibited novel catalytic ability over a broad range of salinity (NaCl 0-3 mol L-1 ). Calcium ion enhanced the proteolytic activity of the enzyme. The Km and Vmax values of the purified protease for casein were calculated to be 4.89 mg mL-1 and 1111.11 U mL-1 , respectively. The protease was strongly inhibited by ethylenediamine tetraacetic acid (EDTA) and phenylmethanesulfonyl fluoride (PMSF). Meanwhile, the protease was stable in the presence of Triton X-100, isopropanol, ethanol or dithio-bis-nitrobenzoic (DTNB), but was inhibited by sodium dodecyl sulfate (SDS), dimethyl sulfoxide (DMSO) or methanol. MALDI -TOF/TOF MS analysis revealed that the protease shared some functional traits with protease produced by Halogranum salarium. Furthermore, it exhibited high hydrolytic activity on silver carp myosin protein. The protease is an alkaline and salt-tolerant enzyme that hydrolyses silver carp myosin with high efficiency. These excellent characteristics make this protease an attractive candidate for industrial use in low-salt fish sauce fermentation. © 2016 Society of Chemical Industry.

Purification and Characterization of a Halotolerant Alkaline Serine Protease from Penicillium citrinum YL-1 Isolated from Traditional Chinese Fish Sauce

ABSTRACTA halotolerant alkaline serine protease from Penicillium citrinum YL-1 which was isolated from traditional Chinese fish sauce was purified by ammonium sulfate precipitation, dialysis, and DEAE 52-Cellulose column, thereby resulting in a 4.66-fold increase in specific activity (110.68 U/mg). The molecular weight (MW) was estimated to be 32.27 kDa using SDS-PAGE analysis. The protease exhibited optimal activity toward the substrate casein at pH 8.0 at 40°C and was stable at pH 6.0–8.0 and 4–30°C. Activity was inhibited by NaCl and retained at 28.3, 21.4 and 18.1% of the initial activity after incubation for 6 h at 20, 25 and 30% NaCl concentrations, respectively. The enzyme was stimulated by Mn2+ and inhibited by K+, Ca2+, Zn2+, Mg2+, Fe2+, and Fe3+. Km and Vmax of the protease for casein were 1.93 mg/ml and 56.81 μg/(min·ml), respectively. Protease activity was strongly inhibited by phenylmethyl sulfonylfluoride (PMSF), which confirmed the serine protease nature of the enzyme. The protease can hydrolyze tilapia protein in the absence or presence of NaCl (5–30%), thus suggesting that this protease is more halotolerant than the protease from other bacteria with high salinity resistance based on the current literature. These properties make the halotolerant alkaline serine protease a suitable candidate enzyme for fish protein hydrolysis during fish sauce fermentation.

Inhibitory Effects of Spices on Biogenic Amine Accumulation during Fish Sauce Fermentation.

The presence of high levels of biogenic amines is detrimental to the quality and safety of fish sauce. This study investigated the effects of ethanol extracts of spices, including garlic, ginger, cinnamon, and star anise extracts, in reducing the accumulation of biogenic amines during fish sauce fermentation. The concentrations of biogenic amines, which include histamine, putrescine, tyramine, and spermidine, all increased during fish sauce fermentation. When compared with the samples without spices, the garlic and star anise extracts significantly reduced these increases. The greatest inhibitory effect was observed for the garlic ethanolic extracts. When compared with controls, the histamine, putrescine, tyramine, and spermidine contents and the overall biogenic amine levels of the garlic extract-treated samples were reduced by 30.49%, 17.65%, 26.03%, 37.20%, and 27.17%, respectively. The garlic, cinnamon, and star anise extracts showed significant inhibitory effects on aerobic bacteria counts. Furthermore, the garlic and star anise extracts showed antimicrobial activity against amine producers. These findings may be helpful for enhancing the safety of fish sauce.

Quantification of viable bacterial starter cultures of Virgibacillus sp. and Tetragenococcus halophilus in fish sauce fermentation by real-time quantitative PCR

Real-time quantitative polymerase chain reaction (qPCR) methods were developed for the quantification of Virgibacillus sp. SK37 and Tetragenococcus halophilus MS33, which were added as starter cultures in fish sauce fermentation. The PCR assays were coupled with propidium monoazide (PMA) treatment of samples to selectively quantify viable cells and integrated with exogenous recombinant Escherichia coli cells to control variabilities in analysis procedures. The qPCR methods showed species-specificity for both Virgibacillus halodenitrificans and T. halophilus as evaluated using 6 reference strains and 28 strains of bacteria isolated from fish sauce fermentation. The qPCR efficiencies were 101.1% for V. halodenitrificans and 90.2% for T. halophilus. The quantification limits of the assays were 103 CFU/mL and 102 CFU/mL in fish sauce samples with linear correlations over 4 Logs for V. halodenitrificans and T. halophilus, respectively. The matrix effect was not observed when evaluated using fish sauce samples fermented for 1–6 months. The developed PMA-qPCR methods were successfully applied to monitor changes of Virgibacillus sp. SK37 and T. halophilus MS33 in a mackerel fish sauce fermentation model where culture-dependent techniques failed to quantify the starter cultures. The results demonstrated the usability of the methods as practical tools for monitoring the starter cultures in fish sauce fermentation.

魚醤油発酵時のヒスタミン蓄積に関わる原因菌の同定および乳酸菌発酵スターター接種によるヒスタミン蓄積抑制効果について

魚醤油発酵時のヒスタミン蓄積に関わる原因菌の同定および乳酸菌発酵スターター接種によるヒスタミン蓄積抑制効果について

Production of a Meat Seasoning Powder Enriched with γ-Aminobutyric Acid (GABA) from Mature Coconut Water Using Pediococcus pentosaceus HN8

The optimal conditions for producing 9.96 g/L GABA (γ-aminobutyric acid) in mature coconut water by Pediococcus pentosaceus HN8 were 10% monosodium glutamate as an additive, an initial pH of 5.92 and 33C under static condition for 24 h. A seasoning meat (112 g/L culture broth), spicy Laab-Numtok powder enriched with 8.2% GABA, was obtained from heating five-time-concentrated culture broth after mixing with other ingredients (table salt, lime juice, fermented fish sauce, chili powder, sliced shallots, ground galangal, sliced kaffir lime leaves and ground roasted rice), and followed by drying. This product met Thai Community Product Standard (493/2547) and it had the ability to inhibit foodborne pathogens. A sensory test showed our product and a popular commercial brand (CB3) had the highest score for all parameters. This work was most useful to add value to the by-product of coconut milk industry by producing a novel meat seasoning powder. Practical Applications Pediococcus pentosaceus HN8 could produce high amounts of γ-aminobutyric acid (GABA) in mature coconut water (a byproduct from coconut milk industry) under static condition for 24 h by optimization on variable factors such as monosodium glutamate, initial pH and incubating temperature. To obtain meat seasoning powder (spicy Laab-Numtok), culture broth enriched with GABA was concentrated by heating, then mixing with ingredients (herbs, salt, fermented fish sauce and ground roasted rice) and finally drying. No significant difference for the sensory test of our product and the most popular commercial brands of spicy Laab-Numtok; however, our product provided sources of GABA, essential amino acids and antibacterial activity for producing safe and healthy Thai top dishes like spicy meats (i.e., chicken, duck and pork). Inexpensive GABA production in this study could be useful for the food industry to produce varieties of the novel meat seasoning products with more benefits to consumers.

Study on microbial and physical changes in fish sauce during fermentation

The production of fermented fish sauce was replicated in the laboratory in order to study the microbiological and physical changes associated with the process. Fish sauce were produced by incubating mixtures of low valued small freshwater fish at different concentrations of sodium chloride (25% as Treatment-I, 30% as Treatment- II and 35% as Treatment-III) for 9 months of fermentation and 15 days of aging. Changes in bacterial composition and physical condition were observed. Due to effect of salt the bacterial population gradually changes and finally salt tolerant bacteria were survived and multiplied. The major bacterial groups identified in fish sauce were Bacillus, Micrococcus, Lactobacillus and Pseudomonas . Due to action of enzymes present in fish body and intestine the solid fish gradually converts in to liquid where salt acts a preservative to prevent the fish from spoilage. Finally the sensory analysis fish sauce was done through panel test. Considering the values of different parameters (salt content, test, colour and flavor) fish sauce produced with 25% salt (Treatment-I) was found to be of best quality.Res. Agric., Livest. Fish.2(2): 375-383, August 2015

Improvement of Fish Sauce Quality by Strain CMC5-3-1: A Novel Species of Staphylococcus sp.

Staphylococcus sp. CMC5-3-1 and CMS5-7-5 isolated from fermented fish sauce at 3 to 7 mo, respectively, showed different characteristics on protein hydrolysis and volatile formation. These Gram-positive cocci were able to grow in up to 15% NaCl with the optimum at 0.5% to 5% NaCl in tryptic soy broth. Based on ribosomal 16S rRNA gene sequences, Staphylococcus sp. CMC5-3-1 and CMS5-7-5 showed 99.0% similarity to that of Staphylococcus piscifermentans JCM 6057(T) , but DNA-DNA relatedness was <30%, indicating that they were likely to be new species. DNA relatedness between these 2 strains was only 65%, suggesting that they also belonged to different species. The α-amino group content of 6-month-old fish sauce inoculated with Staphylococcus sp. CMC5-3-1 was 740.5 mM, which was higher than that inoculated by the strain CMS5-7-5 (662.14 mM, P < 0.05). Histamine was not produced during fermentations with both strains. Fish sauce inoculated with Staphylococcus sp. CMC5-3-1 showed the highest content of total glutamic acid (P < 0.05). The major volatile compound detected in fish sauce inoculated with Staphylococcus sp. CMC5-3-1 was 2-methypropanal, contributing to the desirable dark chocolate note. Staphylococcus sp. CMC5-3-1 could be applied as a starter culture to improve the umami and aroma of fish sauce.

전처리 방법에 따른 홍게(Chionoecetes japonicus) 어간장의 제조 및 품질변화

We explored preprocessing-mediated quality changes in red snow crab fish sauce. A control (C) group and groups treated with autolysis (A), boiling (B), enzymatic hydrolysis (E), and addition of Aspergillus oryzae (K) were formed. The titratable acidity of the K group increased with storage time, whereas that of groups C, A, B, and E decreased. The total and amino nitrogen contents initially increased on storage of all samples, but decreased in later periods. The total plate count (TPC) of the K group was initially 5.26 log CFU/mL and increased to 7.28 log CFU/mL at 3 months of storage. The TPCs of the C, A, B, and E groups were initially <5.00 log CFU/mL and decreased with storage. The lactic acid bacteria count of the K group was initially 4.80 log CFU/mL and increased until month 5 to approximately 6.06 log CFU/mL. The K group scored higher in terms of sensory attributes than the other groups and maintained marketable scores for all relevant properties (color, flavor, off-odor, and overall acceptance). Furthermore, the free amino acid content of the K group was the highest among all groups at approximately 3,000 mg per 100 g. These results suggest that K treatment may be beneficial in the preparation of fermented fish sauce.

시판 액젓 및 간장으로부터 짠맛 증진 물질의 생산 가능성 연구

The quality of commercial fermented fish and soy sauces was evaluated through physicochemical and sensory analysis in order to search a possibility producing salt taste enhancer. Soluble solids (SS) and total nitrogen (TN) content of fish sauces were significantly higher than those of soy sauces (p&lt;0.05). Higher contents of SS and TN resulted in higher levels of free amino acids, degree of hydrolysis, some substances enhancing salt taste, bitterness, and flavor. By GPC chromatogram, nitrogen distribution and salt taste intensity analysis, fraction II of low molecular peptides (220 Da to 1,100 Da), and Lowry nitrogen (soluble nitrogen) that highly correlated salt taste intensity were at the highest levels in salted and fermented anchovy sauce. A desalting process using macroporous adsorption resin (MAR) was not suitable for direct separation from fish sauce due to the loss of low molecular peptides. Salted and fermented anchovy sauce processing method was selected as a method for commercially producing salt taste enhancer because of the highest level of fraction II in spite of higher levels of salinity, bitterness, and long aging time.

Detergent-Stable Salt-Activated Proteinases from Virgibacillus halodenitrificans SK1-3-7 Isolated from Fish Sauce Fermentation.

The NaCl-activated and detergent-stable proteinases from Virgibacillus halodenitrificans SK1-3-7 isolated from fish sauce fermentation were purified and characterized. The enzymes with molecular masses of 20 and 36 kDa showed caseinolytic activity on a zymogram. Optimum azocaseinolytic activity was at 60 °C and pH 9. The proteolytic activity increased in the presence of 10 mM CaCl2 and 0.5 M NaCl and showed high stability at 0-2 M NaCl. The enzymes were stable at pH 4-10 and 10-50 °C. The enzymes preferably hydrolyzed Suc-Ala-Ala-Pro-Phe-pNA and were completely inhibited by phenylmethanesulfonyl fluoride (PMSF), showing subtilisin-like characteristics. Activity and stability remained high in the presence of H2O2 and various surfactants. The enzymes exhibited high stability (>95%) in various organic solvents (DMSO, butanol, ethanol, 2-propanol, and acetonitrile) at concentration of 50%. The V. halodenitrificans SK1-3-7 proteinases showed potential as a biocatalyst in aqueous-organic solvent systems and as an additive in laundry detergent.