ABSTRACT
Salted and fermented seafood (jeotgal) is known for its long shelf life and unique flavor. Despite the existence of various types of salted seafood, the factors influencing this quality have yet to be identified. These factors are essential for improving the quality of salted seafood, optimizing the fermentation process, and advancing the industrialization of fermented foods. Therefore, in this study, we explored microbial dynamics and changes in quality characteristics in three salted seafood items - salted anchovies (MJ), salted cutlass offal (GJ), and salted croakers (HJ), over a 24-month fermentation period. Distinct microbial community profiles, dominated by Tetragenococcus halophilus, Halanaerobium fermentans, and Chromohalobacter canadensis in MJ, GJ, and HJ, respectively, affect the metabolic pathways and the corresponding flavor profiles. The pH of all samples ranged from 5.7-6.0. The titratable acidity was highest in MJ at 1.4% and lowest in HJ at approximately 0.7%. Salinity was below 25% in all samples but slightly lower in MJ. Significant differences were observed in the amino acid, nucleotide, and overall metabolite profiles. MJ exhibited the highest amino acid and nitrogen-related factor levels, such as glutamic acid and hypoxanthine, enhancing flavor complexity. Correlation analysis revealed significant associations among the types, metabolites, and microbial communities. Microbial survival mechanisms in high-salt environments result in the production of unique metabolites, including umami and aroma components as well as precursors of biogenic amines, which can affect the overall quality of the final product. These differences were primarily influenced by the fish type rather than the fermentation time. Our findings provide foundational insights for enhancing fermentation strategies, improving product consistency, and advancing the industrial application of microbial management in seafood fermentation. This study not only fills a significant gap in the current understanding of fermented seafood but also outlines practical approaches for industry applications for the optimization of product quality.
ABSTRACT
Kimchi cabbage, the key ingredient in kimchi, is cultivated year-round to meet high production demands. This study aimed to examine the effects of seasonal harvesting (spring, summer, fall, and winter) on the microbial and metabolic profiles of kimchi during 30 days of fermentation. Lactic acid bacteria distribution is notably influenced by seasonal variations, with Latilactobacillus dominant in fall-harvested kimchi group and Weissella prevailing in spring, summer, and winter. The microbial communities of spring and fall group exhibited similar profiles before fermentation, whereas the microbial communities and metabolic profiles of spring and summer group were similar after 30 days of fermentation. Seasonal disparities in metabolite concentrations, including glutamic acid, serine, and cytosine, persist throughout fermentation. This study provides a comprehensive understanding of the substantial impact of seasonal harvesting of kimchi cabbage on the microbial and metabolic characteristics of kimchi, providing valuable insights into producing kimchi with diverse qualities.
Subject(s)
Brassica , Fermentation , Fermented Foods , Food Microbiology , Seasons , Brassica/microbiology , Brassica/metabolism , Fermented Foods/microbiology , Fermented Foods/analysis , Metabolome , Microbiota , Weissella/metabolismABSTRACT
Mollusca species shell such as oyster shell (OS) and snail shell (SS), are discarded after taking the meat, and the discarded shell causes the environmental problems. Therefore, recycling shell waste could potentially eliminate the environmental problems. This study aimed to evaluate the potential of OS and SS as natural calcium resources. The minerals, calcium, magnesium, potassium, phosphorus and sodium were analyzed in OS and SS extracts. Among them, the calcium content was the highest: 36.87 (%) and 33.42 (%) in the OS and SS extracts, respectively. Further, the content of ionized bioavailable form of calcium in OS and SS was higher than that of CaCO3 under simulated gastrointestinal digestion conditions. Additionally, OS and SS were added to kimchi, and their inhibitory effect on kimchi acidification was evaluated by assessing pH, titratable acidity and microbial analysis. As the results indicated that the addition of OS and SS had little effect on inhibiting the growth of lactic acid bacteria. However, it was confirmed that calcium neutralizes the organic acids produced during fermentation. Overall, the results of this study provide preliminary information on the re-use of OS and SS extracts as ionized natural calcium supplements and fermentation retardants.
ABSTRACT
The cultivar of red pepper used in kimchi contributes to spiciness, red color, and fermentation characteristics. Capsaicinoids are the main components of red pepper. Therefore, understanding changes in metabolites during kimchi fermentation according to capsaicinoid concentration is necessary to control the quality of kimchi. The purpose of this study was to investigate the effect of capsaicinoids on metabolites during kimchi fermentation. To profile the effect of capsaicinoid concentrations on kimchi fermentation, five kimchi samples were prepared using different concentrations of capsaicinoids (4, 12, 30.7, 40.9, and 50.3 mg/kg) and stored at 4 °C for 28 days. During kimchi fermentation, pH, titratable acidity, capsaicinoid concentration, total viable and lactic acid bacteria, free sugars, amino acids, and microbial community were evaluated. Each result was statistically analyzed for changes in capsaicin concentration and fermentation time. The capsaicinoid concentration did not change during kimchi fermentation but the growth of lactic acid bacteria changed. According to the growth of lactic acid bacteria, free sugar, amino acids, and microbial community changed with the capsaicinoid concentration. Overall, the results of this study provide preliminary information on the use of red pepper and capsaicinoids in the kimchi industry.
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Garlic, a key ingredient in kimchi, is an indispensable source of lactic acid bacteria, which are essential for fermentation. This study explored the effects of various garlic varieties on kimchi fermentation, focusing on changes in microbial communities and metabolite profiles. We observed that the type of garlic used did not significantly alter the microbial community. However, the presence of garlic itself made a significant difference. Specifically, kimchi with garlic showed higher abundance of Leuconostoc and Weissella, which are bacteria primarily responsible for kimchi fermentation. Additionally, kimchi containing garlic had increased levels of mannitol and fructose, which significantly influence taste; however, lactic acid and putrescine levels were decreased. Therefore, the addition of garlic directly contributes to the flavor profile of kimchi. Sixty-two metabolites were identified using gas chromatography and mass spectrometry. The variety of garlic added influenced the metabolite profiles of kimchi, particularly in the later stages of fermentation. These profiles were categorized based on the garlic's origin, whether from southern or northern ecotypes (R2X = 0.933, R2Y = 0.986, Q2 = 0.878). These findings confirm that both the presence and the variety of garlic significantly impact the microbial ecology and metabolites during kimchi fermentation, underscoring its essential role in the process.
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Fermentation of salted shrimp involves the interaction of multiple factors. However, studies of the effects of shrimp variety and fermentation temperature on metabolites generated during fermentation are limited. Therefore, we investigated the effects of the shrimp variety, fermentation temperature, and fermentation period on the composition of fermented salted shrimp. Four different varieties of salted shrimp, namely, Detteugijeot (SSA), Red shrimp jeot (SSB), Chujeot (SSC), and Yukjeot (SSD), were prepared and stored at 5 and 10 °C for 5 months. The pH values ranged from 6.71 to 6.99, with SSD showing the lowest pH at both temperatures. Although total nitrogen content remained relatively constant, amino nitrogen exhibited an upward trend after 2 months and was particularly increased at 10 °C. This increase was attributed to variations in microorganisms and enzymes in the salted shrimp. Except for proline, citrulline, and ornithine, amino acid levels increased during fermentation with the highest amounts detected in SSA. Additionally, the levels of glutamic acid and branched-chain amino acids were found to be sensitive to fermentation temperature. Amino acid levels were apparently affected by species-specific metabolic pathways of the microorganisms present in each salted shrimp. Compared to the other varieties, SSB had significantly higher contents of adenosine triphosphate and hypoxanthine. A high hypoxanthine content could contribute to increased bitterness and an umami taste profile. Furthermore, the correlation between salted shrimp and metabolites was unique in SSB, whereas partial clustering was observed between the SSA and SSC.
ABSTRACT
Gelatinized starch sauce, one of the sub-ingredients have been widely used in kimchi for their roles in increasing viscosity of kimchi seasoning, and fermentation. Gelatinized glutinous rice (GGR), which is one of the most used starch sources in kimchi preparation. However, GGR is accelerated to the fermentation process but lead to a reduction in the shelf life of the kimchi. Therefore, in this study, we demonstrate the effectiveness of using maltodextrin (MD) as a novel starch source instead of GGR to slow down the rate of kimchi fermentation. The properties of the kimchi with MD and GGR fermentation (free sugar content, organic acid content, pH, and acidity) as well as their microbial growth rates after 12 days of fermentation were compared. After fermentation of 12 days, the free sugar of GGR-kimchi (GGRK) increased more rapidly than those of MD-kimchi (MDK), while higher sugar alcohol (mannitol) and organic acid contents were observed for GGRK than for MDK. Furthermore, initial aerobic and lactic acid bacteria counts were higher for GGRK than for MDK. These results indicate that fermentation proceeds at a slower rate in MDK than in GGRK, and they will provide a basis for further research into storage of kimchi. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-023-05742-y.
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Salt is an essential ingredient in the kimchi fermentation process. Solar salt has antioxidant, anti-cancer, and anti-obesity properties. The aim of this study was to determine the antioxidant and anti-inflammatory effects of solar salt brined kimchi. Purified salt (PS), dehydrated solar salt (DSS), 1-year aged solar salt (SS1), and 3-years aged solar salt (SS3) were investigated. Anti-inflammatory effects were determined by analyzing cytotoxicity, nitric oxide (NO) production, and inflammation-related gene expression in lipopolysaccharide-treated RAW264.7 cells. Antioxidant activities of DSS, SS1, and SS3 were higher than that of PS. Solar salt significantly inhibited NO production with low cytotoxicity and decreased inflammation-related gene expression. Kimchi containing solar salt (DSSK, SS1K, and SS3K) showed higher antioxidant activity than PSK. Additionally, DSSK, SS1K, and SS3K significantly inhibited NO production and decreased the expression of inflammation-related genes. Owing to the antioxidant and anti-inflammatory effects, using solar salt in kimchi preparation could have potential health benefits.
ABSTRACT
It is important to optimize the drying process, along with the concentration of drying aid and the inlet air temperature, in order to obtain products with better physicochemical properties. Onion-stevia leaf hot extract powders were prepared using gum arabic (GA) and whey protein concentrate (WPC). Inlet air temperature and carrier concentrations were optimized using response surface methodology. The drying yield of powdered extracts was 14.39-74.32%, L*- value was 52.66-66.98, bulk density was 0.36-0.75 (g/cm3), moisture content was 2.40-11.57%, water solubility index was 30.32%-97.46%, and mean particle size D[4,3] was 9.13-88.01 (µm). For both GA- and WPC-based powders, optimal inlet air temperatures and carrier concentrations were 148.81 and 144.62°C, and 11.58 and 12.03% (w/v), respectively. GA powders had a higher glass transition temperature (76.49°C) as compared to WPC powders (48.12°C) or maltodextrin as control (55.49°C). Sweetness (5.0/7.0) and overall acceptability (4.3/7.0) scores were higher for GA powders as compared to WPC powders (3.7/7.0 and 3.4/7.0), respectively. Conclusively, GA is a better carrier than whey protein for preparing spray-dried onion-stevia powder that can be used as a natural sweetener.
ABSTRACT
During storage and fermentation of kimchi, white film-forming yeasts (WFY) are generated on the surface of kimchi under various conditions. These yeasts include Candida sake, Pichia kudriavzevii, Kazachstania servazzii, Debaryomyces hansenii, and Hanseniaspora uvarum. Because of the off-odor and texture-softening properties of WFY that degrade the quality of kimchi, a method to prevent WFY is required. In this study, cinnamaldehyde (CIN) and gold nanoparticles (AuNPs) with a large surface area were grafted on a paper surface, which was termed the "Paper_AuNPs_CIN" film. CIN is an antimicrobial agent that is approved for use in food applications. In the as-fabricated Paper_AuNPs_CIN film, antimicrobial CIN molecules were physically adsorbed to the surface of AuNPs and simultaneously chemically synthesized on the paper surface via the imine reaction. The Paper_AuNPs_CIN film exhibited greater antimicrobial activity against the three WFY strains than a Paper_CIN film (which contains only CIN molecules). Since more CIN molecules were adsorbed to the large surface area of the paper-reduced AuNPs, the Paper_AuNPs_CIN film exhibited a higher antimicrobial activity. Using AuNPs and CIN simultaneously to inhibit the growth of WFY is a novel approach that has not yet been reported. The morphology and elemental mapping of the functionalized films were examined via scanning electron microscopy and energy-dispersive spectroscopy, elemental composition was analyzed via inductively coupled plasma optical emission spectroscopy, and chemical bonding and optical properties were investigated via Fourier transform infrared spectroscopy and diffuse reflectance spectroscopy. Additionally, agar-well diffusion assays were used to determine the antimicrobial activity against three representative WFY strains: C. sake, P. kudriavzevii, and K. servazzii.
ABSTRACT
The effect of salt concentration (3%, 6%, and 9%) on the mass transfer kinetics of Kimchi cabbage during osmotic dehydration was investigated, including its influence on textural and microstructural properties and salt distribution. First, kinetics was analyzed using diffusion theory to determine the impact of the factors on moisture and salt transfer. Subsequently, using the Peleg, Azuara, Henderson-Pabis, and Page models, mathematical modeling of mass transfer (water loss and salt gain) was investigated. According to the statistical analysis, the Peleg model provided the best fit for the experimental results under the operating conditions. In addition, a novel viewpoint was proposed in which the salt content of Kimchi cabbage may be indirectly forecasted by monitoring solution salinity during osmotic dehydration. Higher salt concentration resulted in decreased hardness, gumminess, and chewiness in Kimchi cabbage. Scanning electron microscopy and energy-dispersive X-ray mapping images showed an intensification of moisture and salt transport with increasing salt content, which were confirmed using modeling studies. The results could be applied in the prediction of the target salinity of Kimchi cabbage during the salting process and could facilitate the improvement of final Kimchi product quality by producing salted Kimchi cabbage with uniform salinity.
Subject(s)
Brassica rapa , Brassica , Fermented Foods , Brassica/chemistry , Kinetics , Dehydration , Sodium Chloride , Sodium Chloride, DietaryABSTRACT
The taste of kimchi is greatly affected by the salt type used during fermentation. Here, we investigated the effects of salts with different mineral contents on the microbial community and metabolite profiles of fermented kimchi using multivariate statistical analysis. We evaluated different types of salt used to prepare kimchi, namely, solar salt aged for 1 year, solar salt aged for 3 years, dehydrated solar salt, and purified salt. The main microorganisms detected in kimchi were Weissella koreensis, Leuconostoc mesenteroides, and Latilactobacillus sakei. Leuconostoc and Weissella were mainly present in kimchi supplemented with solar salt. However, a high proportion of L. sakei was present in kimchi supplemented with purified salt and dehydrated salt. Additionally, using GC-MS-based metabolite analysis, we revealed that the content of free sugars, organic acids, and amino acids differed in kimchi fermented with different salt types. Therefore, we demonstrated that salt type had a pronounced effect on the resultant microbial community and the type and concentration of metabolites present in fermented kimchi.
ABSTRACT
BACKGROUND: To isolate polysaccharides with enhanced immunostimulatory activity from Dendrobium officinale, which is used as a herbal medicine in China and Southeast Asia, D. officinale (DO) was pretreated with organic solvents (DOOS) or puffing at 7.5 and 9.0 kgf (7.5DO and 9DO). Hot-water extracts (DOOS-HW, 7.5DO-HW and 9DO-HW) were prepared from each pretreated DO, along with non-pretreated DO, and crude polysaccharides (DO-CP, DOOS-CP, 7.5DO-CP and 9DO-CP) were fractionated from each hot-water extract using ethanol (five volumes). RESULTS: When their immunostimulatory activities were compared by macrophage stimulation and intestinal immune system modulation via Peyer's patches, DOOS-CP showed more potent activity than DO-CP. However, crude polysaccharides fractionated from puffed DO showed significantly lower activity than non-puffed DO and DOOS. The most active polysaccharide contained 95% or more neutral sugar, and the composition ratio of mannose and glucose was 3.0, whereas the lowest polysaccharide content was 2.0 or less. In addition, DOOS-CP was a somewhat refined fraction containing a major peak, representing a molecular weight of 250 kDa, despite being a crude polysaccharide. CONCLUSION: These results suggest that pretreatment of D. officinale with organic solvents may enhance the immunostimulatory activity of polysaccharides and affect the mannose/glucose ratio of polysaccharides, which plays an important role in immunostimulation. © 2021 Society of Chemical Industry.
Subject(s)
Dendrobium , Dendrobium/chemistry , Glucose , Mannose , Plant Extracts/chemistry , Polysaccharides/chemistry , Solvents , Sugars , WaterABSTRACT
Effect of capsaicinoids in hot pepper powder (HP) contains various chemical compounds, including capsaicin and dihydrocapsaicin, which are the main ingredients of the spicy taste. To evaluate the effect of HP on the microbial community in kimchi fermentation, kimchi [kimchi-HP, kimchi-HPE and kimchi-HPER made by adding HP, HP alcohol extract (HPE) and HPE residues (HPER)] was fermented at 4°C for 28 days. The pH and titratable acidity of the samples and the number of bacteria changed with fermentation time. Kimchi-HPER had significantly higher total viable and lactic acid bacteria (LAB) than other samples after 28 days of fermentation. The capsaicinoids content did not differ before and after fermentation, whereas the major free sugar content decreased, and the mannitol content increased. The principal component analysis (PCA) biplots showed similar patterns between kimchi-HP and -HPE. It was confirmed that Leuconostoc and Weissella were related to the initial fermentation, and Lactobacillus was involved in late fermentation. Kimchi-HP and kimchi-HPE increased the ratio of Lactobacillus sakei and decreased that of Leuconostoc mesenteroides compared to kimchi-HPER. Overall, these results revealed that capsaicinoids contained in HP affected Lactobacillus proliferation and mannitol increase during kimchi fermentation.
Subject(s)
Bacteria/drug effects , Brassica rapa/microbiology , Capsaicin/analogs & derivatives , Capsaicin/pharmacology , Capsicum/chemistry , Fermented Foods/analysis , Sugars/analysis , Animals , Bacteria/growth & development , Bacteria/isolation & purification , Fermentation , Food Microbiology , Powders/chemistryABSTRACT
Changes in color, browning indices, enzyme activity, and physical and chemical quality during the storage period were investigated to assess the effectiveness of storage period extension along with the addition of ascorbic acid (AA) and citric acid (CA) to salted Chinese cabbage. After 16 days of storage, the change in chromaticity value showed treatment with 0.5% CA showed the lowest change in the brown index during the storage period. The control showed the highest residual activity of polyphenol oxidase among control, AA, and CA-treated salted cabbage. AA and CA treatment effectively inhibited the initial populations of microorganisms including total aerobic bacteria, lactic acid bacteria, and yeast and molds in salted Chinese cabbage during storage. Further, the texture, i.e., hardness, chewability, and elasticity, tended to decrease with increasing storage. These results suggest that treatment with AA could help maintain the quality of salted Chinese cabbage during the storage period.
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A case of Taenia asiatica infection detected by small bowel series and colonoscopy is described. The patient was a 42-year-old Korean man accompanied by discharge of movable proglottids via anus. He used to eat raw pig liver but seldom ate beef. Small bowel series radiologic examinations showed flat tape-like filling defects on the ileum. By colonoscopy, a moving flat tapeworm was observed from the terminal ileum to the ascending colon. The tapeworm was identified as T. asiatica by mitochondrial DNA sequencing. The patient was prescribed with a single oral dose (16 mg/kg) of praziquantel.
Subject(s)
Colonoscopy , Taenia/isolation & purification , Taeniasis/diagnosis , Animals , Base Sequence , Colon/parasitology , Colon/pathology , DNA, Helminth/chemistry , DNA, Helminth/genetics , DNA, Mitochondrial/chemistry , DNA, Mitochondrial/genetics , Feeding Behavior , Humans , Ileum/diagnostic imaging , Ileum/parasitology , Ileum/pathology , Korea , Male , Molecular Sequence Data , Radiography, Abdominal , Sequence Alignment , Sequence Analysis, DNA , Taenia/classification , Taenia/genetics , Taeniasis/parasitologyABSTRACT
After publishing results of a study that revealed diarrheagenic and emetic activity in 4-5-day old mice infected with Kudoa septempunctata (K. septempunctata) spores, the Korea Centers for Disease Control and Prevention reported 11 events of "Kudoa food poisoning" in 2015. The epidemiological design of the previous study was descriptive rather than analytical; therefore, this study aimed to further investigate the pathogenicity of K. septempunctata. Academic articles showing evidence of the pathogenicity of K. septempunctata were searched via PubMed using the citation discovery tool. Information regarding the kinds of experimental animals and inoculum spores used, as well as study results were extracted. Four articles evaluating the pathogenicity of Myxospran parasites were selected; the first article suggested the pathogenicity of K. septempunctata, while the remaining three articles reported no abnormal symptoms or histopathologic changes. Our findings indicate that there is weak evidence supporting the pathogenicity of K. septempunctata. Further studies evaluating the pathogenicity of K. septempunctata are needed urgently.
Subject(s)
Disease Outbreaks , Fish Diseases/parasitology , Flounder/parasitology , Foodborne Diseases/etiology , Intestinal Diseases, Parasitic/parasitology , Myxozoa/pathogenicity , Parasitic Diseases, Animal/parasitology , Animals , China , Foodborne Diseases/epidemiology , Foodborne Diseases/parasitology , Humans , Intestinal Diseases, Parasitic/epidemiology , Japan , Mice, Inbred Strains , Republic of Korea/epidemiology , Seafood/parasitology , Spores , VirulenceABSTRACT
The quality characteristics of pork loin frozen by cryogenic immersion were examined, such as the drip loss, cooking loss, water holding capacity, moisture content, protein solubility, lipid and protein oxidation, color, and microstructure, and compared after different tempering methods: radio frequency (27.12MHz), water immersion, forced-air convection, and microwave tempering. Forced-air tempering was the most time-consuming process, whereas electromagnetic energy methods (radio frequency and microwave) were the shortest. The tempering rate of radio frequency at 400W was 5 and 94 times greater than that obtained with water immersion and forced-air tempering, respectively. The drip loss, water holding capacity, moisture content, color, and microstructure of pork samples all declined as a result of microwave tempering. By contrast, the least degree of changes in the drip loss, microstructure, and color of the pork loin samples was obtained with radio frequency tempering, suggesting its potential application in providing rapid defrosting without quality deterioration in the frozen meat industry.
Subject(s)
Food Handling , Food Quality , Freezing , Red Meat , Animals , Color , Dietary Proteins/analysis , Muscle, Skeletal/chemistry , Swine , Thiobarbituric Acid Reactive Substances/analysis , Volatile Organic Compounds/analysis , Water/analysisABSTRACT
PURPOSE: Recent studies have used the term "gastroallergic anisakiasis" to describe incidental gastrointestinal infection with Anisakis spp. larvae, proposed as a causative agent of food hypersensitivity. However, it is unknown whether this condition represents an independent disease entity distinguishable from acute gastric anisakiasis. To better understand the role of the allergic response in Anisakis infections we examined the clinical and immunological implications of Anisakis-specific IgE. METHODS: A prospective study was performed in a geographic region where the consumption of raw seafood is common. Case subjects who had been clinically diagnosed with gastroallergic anisakiasis were selected, along with controls who frequently ate raw seafood but had never experienced gastroallergic anisakiasis-like symptoms. Clinical and immunological features were compared based on atopic status, sensitization rates to Anisakis, and serum titer of Anisakis-specific IgE. RESULTS: Seventeen case subjects and 135 controls were included in this study. The case subjects had experienced gastrointestinal symptoms after raw seafood ingestion, along with additional mucocutaneous, respiratory, or multisystemic symptoms. Case subjects were significantly sensitized to Anisakis excretory-secretory product and crude extract compared with controls (76.5% vs 19.3%, P<0.001, and 88.2% vs 30.3%, P<0.001, respectively). Anisakis-specific serum IgE titers were also significantly higher in case subjects than in controls. Both the results of skin prick tests and elevated Anisakis-specific IgE titers (>17.5 kU/L) were found to be reliable indicators for the diagnosis of gastroallergic anisakiasis. CONCLUSIONS: Among patients presenting acute gastric anisakiasis-like symptoms, a diagnosis of gastroallergic anisakiasis may be strongly supported by a high Anisakis-specific IgE titer.
ABSTRACT
Several studies have reported that the citrus red mites Panonychus citri were an important allergen of citrus-cultivating farmers in Jeju Island. The aim of the present study was to purify and assess properties of a cysteine protease from the mites acting as a potentially pathogenic factor to citrus-cultivating farmers. A cysteine protease was purified using column chromatography of Mono Q anion exchanger and Superdex 200 HR gel filtration. It was estimated to be 46 kDa by gel filtration column chromatography and consisted of 2 polypeptides, at least. Cysteine protease inhibitors, such as trans poxy-succinyl-L-leucyl-amido (4-guanidino) butane (E-64) and iodoacetic acid (IAA) totally inhibited the enzyme activities, whereas serine or metalloprotease inhibitors did not affect the activities. In addition, the purified enzyme degraded human IgG, collagen, and fibronectin, but not egg albumin. From these results, the cysteine protease of the mites might be involved in the pathogenesis such as tissue destruction and penetration instead of nutrient digestion.