Recently, active research in Korea and worldwide has begun to focus on gene function and cultivar development using gene editing tools. This research, in addition to studies on edible mushroom, aims to enhance the physical and biochemical characteristics of mushrooms for applications in materials and substance production. For these studies, efficient isolation of protoplasts from the target mushroom is critical. However, several commercial cell wall-lysing enzyme cocktails, including Novozyme234, Glucanex, and Lysing enzymes, have recently been discontinued. In this study, we aimed to identify alternative enzyme systems to replace the discontinued cell wall-lysing enzymes for stable isolation of protoplasts from Ganoderma lucidum. To select an optimal osmotic buffer, enzyme function in 0.6 and 1.2 M Sorbitol, Sucrose, Mannitol, and KCl was assessed. The effect of reaction time was also evaluated. Protoplast isolation efficiency of each alternative enzyme was tested using lysing enzymes from Trichoderma harzianum, Chimax-N, and Yatalase, either individually or in combination. This matrix of studies identified enzymes and optimal conditions that could replace the discontinued lysing enzymes.

A new fumigant, carbonyl sulfide (COS), has potential for use as a replacement for methyl bromide, yet its mechanism of toxicity to insects remains poorly understood. In this study, transcriptome analysis was performed on Tribolium castaneum malpighian tubules and fat bodies, which are known to play an essential role in energy storage and utilization in insect species. In total, upon exposure to COS, 3,034 and 2,973 genes were differentially expressed in the T. castaneum malpighian tubules and fat body, respectively. These differentially expressed genes comprise a significant number of detoxification-related genes, including 105 P450s, 18 glutathione S-transferases (GSTs), 82 ABC transporters, 25 UDP-glucosyltransferases and 42 carboxylesterases and mitochondrial–related genes, including 9 complex Ⅰ genes, 2 complex Ⅱ genes, 1 complex Ⅲ gene, 9 complex IV genes, 8 complex V genes from both malpighian tubules and fat body tissues. Moreover, KEGG analysis demonstrated that the upregulated genes were enriched in xenobiotic metabolism by ABC transporters and drug metabolism by other enzymes. We also investigated the role of carbonic anhydrases (CAs) in toxicity of COS using dsRNA treatment in T. castaneum. These results show that CA genes have a key role in toxicity of the COS. Furthermore, the results of transcriptomic analysis provide new insights into the insecticidal mechanism of COS fumigation against T. castaneum and eventually contribute to the management of this important stored grain pests.

본 연구는 아그니와 효소의 종류와 기능을 고찰하고 활성화를 위한 섭생 요법에 대해 소개하였다. 음식 소화에 있어서 아그니와 효소의 불균형은 인체의 신진대사에 악영향을 주어 질병에 노출을 증 가시킨다. 섭생 요법은 소화와 대사를 촉진하고, 독소를 제거하고, 면역력을 높이는 기초이며 건강하 기 위한 중요한 요법이다. 아유르베다 의학의 섭생 요법은 개인의 도샤(doṣa)에 따라 다른 효과를 나 타낼 수 있으며, 일상생활의 규칙성, 식사 시간이나 양, 조리방법 등을 고려해 도샤에 맞는 음식을 섭 취해야 한다. 현대 의학 효소는 화학적인 분자이며 과학적인 근거를 바탕으로 수치화되고 정밀화되고 있다. 효소 활성화를 통해 음식의 칼로리와 영양소가 인체에 흡수되는 정도를 파악할 수 있고 다양한 조리방법으로 흡수 효율을 높이기 위해 노력하고 있다. 또한 세계보건기구(WHO)는 건강한 식사와 생 활 방식을 통해 만성 질환과 감염병을 예방하고 건강을 증진하기 위해 섭생 요법을 권장하고 있다. 이 논문을 통해 우리는 아유르베다 의학과 현대 의학이 공통적으로 강조하고 있는 섭생 요법이 건강 을 유지하는 기본임을 알 수 있을 것이다.

The present studies explored the possibility of immobilizing phosphocholine (PC) liposomes on the surface of graphene oxide (GO) which was pre-adsorbed with two kinds of enzymes, horseradish peroxidase and glucose oxidase. The transmission electron microscope images showed that the PC liposomes adsorbed onto the GO surface kept integrity. By using 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS)-encapsulated liposomes, a one-step colorimetric assay for glucose was developed. In the presence of glucose, the GO nanocomposites catalyzed the cascade enzymatic reaction producing colorimetric signals directly. Under the optimal conditions, the GO nanocomposites produced linearly increased colorimetric signal with increased concentrations of glucose ranging from 50 to 500 μM. The detection limit was 33 μM. The GO nanocomposites also exhibited good selectivity for the detection of glucose and were able to detect glucose in human serum.

최근 살진균제는 세계 식량 안보에 없어서는 안될 필수 요소이며, 그 사용량은 증가하고 있다. 살진균제는 직접적 또는 간접적으로 곤충에 영 향을 미쳐 유전자 및 분자 수준의 변화를 일으킨다. 곤충은 다양한 해독 매커니즘을 통해 살진균제를 포함한 농약으로부터 유발되는 활성산소 (ROS) 독성을 제거한다. 본 연구는 살진균제 캡탄의 비치명적 투여량(0.2, 2, and 20 μg/μL)을 주입 후 갈색거저리의 유충에서 해독효소의 mRNA 발현량을 분석했다. 갈색거저리의 전사체 분석을 통해 해독 매커니즘 관련 유전자인 퍼옥시다제(POX), 카탈라제(CAT), 슈퍼옥사이드 디스뮤타제(SOD) 및 글루타티온-S-트랜스퍼라제(GST)를 발굴하였다. 처리 24시간 후 TmPOX5 mRNA가 유의하게 증가한 것으로 나타났다. 처리 3 시간 후 TmSOD4의 mRNA가 유사하게 증가하였다. 또한 2 μg/μL 처리 24시간 후 TmCAT2의 mRNA 가 유의하게 증가하였다. 캡탄 노출 후 TmGST1 및 TmGST3의 mRNA 발현량도 증가하였다. 결론적으로, TmPOX5 및 TmSOD4 유전자는 갈색거저리에서 캡탄 노출에 대한 바이오마커 또는 생체이물 센서로 작용할 수 있음을 시사한다.

This in vitro study investigated the enhancement of rumen bacterial adhesion on a substrate to increase the digestibility of rice straw in Hanwoo cattle. The rice straw was pretreated with enzymes, probiotics, or ammonia, and the effects on the enhancement of bacterial adhesion and fiber degradation were analyzed using in vitro rumen fermentation. Enzyme treatment included spraying of cellulase and xylanase at 40 and 120 U per g of rice straw, respectively; Saccharomyces cerevisiae culture of 1.0×107 CFU was sprayed as a probiotic treatment per gram of rice straw; ammonia was sprayed at 0.3% per gram of rice straw. Following enzyme treatment, Fibrobacter succinogenes formed a higher adhesion colony than the control group (7.26±0.03 and 8.43±0.20) after 6h and 12h of in vitro culture (p<0.05), respectively. Attachment of Ruminococcus flavefaciens also increased following enzyme treatment (p<0.05) after 6 and 12 h compared to that of the control (5.18±0.06 and 6.60±0.15); and R. albus attachment showed a significant increase compared to that of the control (5.94±0.15) after 6 h of incubation (p<0.05). Probiotic treatment increased attachment of F. succinogenes in comparison with untreated rice straw after 6 h and 12 h of fermentation (p<0.05); R. flavefaciens attachment showed an increase only after 6 h of culture (p<0.05); R. albus was not affected. Attachment of F. succinogenes, R. flavefaciens, and R. albus increased with ammonia treatment after 6 h and 12 h (p<0.05). Dry matter digestibility was higher after the enzyme treatment (3.45±0.21 and 7.04±0.09) than in the control group(1.85±0.08 and 3.94±0.04) after 6 and 12 h of in vitro culture (p<0.05), respectively. It was also higher than that of untreated rice straw after probiotic and ammonia treatments (p<0.05). There was an increase in the enhancement levels of bacterial adhesion depending on the type of fibrolytic bacteria following enzyme, probiotic, and ammonia treatment. These treatments improved digestibility. This enhancement is considered to be greater following enzyme and ammonia treatments than with probiotics.

본 연구에서는 국내 다빈도로 사용되고 있는 가공보조제 효소제 7종에 대한 최대 사용량 기반 세포독성을 연구하였다. 기원이나 원산지 및 제조형태가 다른 효소제가 시판되고 있는 경우 모두 시험대상에 포함하였고, 효소제의 경우 가공과정에 사용 후 최종제품에서 잔존하지 않거나 불활성화되기 때문에 native 및 inactivated 두 형태에 대한 세포독성을 확인하였다. 세포처리 최고 농도는 품목 제조보고서를 기반으로 다빈도 첨가 식품품목 및 그 최대 사용량과 국민영양통계 일일섭취량을 기반으로 설정하였 다. 그 결과, native 및 inactivated 효소제 모두 세포성장 및 세포막 손상에 의한 세포사멸을 유발하지 않는 것으로 나타났다. Native 효소제의 경우 종국 3종 모두에서 ROS 유발이 확인되었으나, inactivated 종국에서 ROS의 유의적 증가는 나타나지 않았다. 일부 native 형태의 종국 및 프로테아제에서 β-hexosaminidase 용출에 의한 탈 과립화가 유발될 수 있는 것으로 분석되었으나, 모든 inactivated 효소 제에서 탈 과립화는 유의적으로 나타나지 않았다. 이러한 결과는 효소제의 기원에 따라서 native 형태의 경우 ROS 유발에 의한 산화스트레스 및 탈 과립화와 같은 알레르기 원성 세포독성을 나타낼 수 있으나, 실제 가공제품에 잔존 하는 불활성화된 효소제에 의한 세포독성 영향은 낮음을 제시한다 할 수 있다. 이와 같은 연구결과는 안전성 정보 가 미흡한 가공보조제 효소제에 대한 구체적인 독성자료 및 향후 in vivo 독성연구를 위한 기초자료로 활용될 수 있을 것이다.

In marine ecosystems, the biosynthesis and catabolism of dimethylsulfoniopropionate (DMSP) by marine bacteria is critical to microbial survival and the ocean food chain. Furthermore, these processes also influence sulfur recycling and climate change. Recent studies using emerging genome sequencing data and extensive bioinformatics analysis have enabled us to identify new DMSP-related genes. Currently, seven bacterial DMSP lyases (DddD, DddP, DddY, DddK, DddL, DddQ and DddW), two acrylate degrading enzymes (DddA and DddC), and four demethylases (DmdA, DmdB, DmdC, and DmdD) have been identified and characterized in diverse marine bacteria. In this review, we focus on the biochemical properties of DMSP cleavage enzymes with special attention to DddD, DddA, and DddC pathways. These three enzymes function in the production of acetyl coenzyme A (CoA) and CO2 from DMSP. DddD is a DMSP lyase that converts DMSP to 3-hydroxypropionate with the release of dimethylsulfide. 3-Hydroxypropionate is then converted to malonate semialdehyde by DddA, an alcohol dehydrogenase. Then, DddC transforms malonate semialdehyde to acetyl-CoA and CO2 gas. DddC is a putative methylmalonate semialdehyde dehydrogenase that requires nicotinamide adenine dinucleotide and CoA cofactors. Here we review recent insights into the structural characteristics of these enzymes and the molecular events of DMSP degradation.

뽕잎 (Morus alba L.)을 노루궁뎅이 버섯균사체(Hericium erinaceum)로 발효시켜 열수 추출한 발효물(MA-HE)이 항산화 효소인 Cu,Zn-superoxide dismutase (SOD)와 ceruloplasmin (CP)의 산화적 변형에 미치는 영향을 관찰하였다. 세포질에서 중요한 항산화 활성을 나타내는 Cu,Zn-SOD와 CP는 모두 구리이온을 포함하는 단백질이다. MA-HE은 peroxyl radical에 의해 변형되는 Cu,Zn-SOD와 CP를 모두 보호하였다. MA-HE은 peroxyl radical에 의한 Cu,Zn-SOD의 단백질 fragmentation과 효소의 inactivation을 모두 효과적으로 억제하였다. 또한, MA-HE는 CP의 fragmentation과 inactivation도 효과적으로 억제하였다. MA-HE가 peroxyl radical을 소거할 수 있는 기능을 가지고 있는 지를 확인하기 위해 thiobarbituric acid를 이용한 활성 측정법으로 관찰한 결과, MA-HE는 농도가 증가함에 따라 비례적으로 peroxyl radical 소거 활성이 높아지는 것으로 관찰되었다. MA-HE는 100 μg/mL에 서 44.03% 소거활성을 나타냈다. 따라서, MA-HE는 peroxyl radical을 효과적으로 소거함으로서 세포 내의 항산화 효소를 산화적 손상으로부터 보호할 것으로 생각된다. 이와 같은 결과는 MA-HE가 세포 내에서 발생하는 활성산소종을 효과적으로 소거하여 산화적 스트레스에 의해 야기되는 세포 독성에 대한 보호 작용을 할 것으로 생각된다.

This study investigated the bread-making properties of rice bread supplemented with HPMC (hydroxypropyl methylcellulose) 1~3% and gluzyme (glucose oxidase), fungamyl (fungal α-amylase) and pentopan (xylanase+ hemicellulases) (0.0015~0.0090%). The viscoelastic properties of the dough with HPMC 1~3% were analyzed. When the rice flour was supplemented with HPMC 1~3%, the viscoelastic properties of the dough tended to increase as the amount of added HPMC was increased. The physicochemical characteristics of the rice bread with HPMC, gluzyme, fungamyl, and pentopan were analyzed. Supplementing the rice flour with HPMC, gluzyme, fungamyl, and pentopan had a significant effect on the volume (p<0.01) and specific volume (p<0.001) of the rice bread. Supplementing the rice flour with 3% HPMC and 0.0045% or 0.0090% pentopan had a significant effect on increasing the volume (p<0.01) and specific volume (p<0.001) of the rice bread. Supplementing the rice flour with 3% HPMC, 0.0023% gluzyme and 0.0015% fungamyl had a significant effect on increasing the volume (p<0.01) and specific volume (p<0.001) of the rice bread. These results suggest that supplementing the rice flour with HPMC, gluzyme, fungamyl and pentopan is effective for the production of rice bread.

The baking properties of rice bread with polydextrose (3, 6, and 9%), enzymes (0.006, 0.009, and 0.018%) and egg (1.32 and 2.64%) were investigated. The specific gravity and color (L, a, b) of the dough, as well as the appearance, color (L, a, b) and texture of the rice bread were analyzed. The springiness, chewiness, gumminess (p<0.01) and hardness (p<0.001) of the rice bread tended to increase as the amount of added polydextrose increased. Replacement of rice flour with hemicellulase, glucose oxidase and fungal amylase are effective for producing rice bread. Replacement of rice flour with 0.009% glucose oxidase and 0.006% fungal amylase had a significant effect on increasing the volume and decreasing the hardness of the rice bread (p<0.001). Replacement of rice flour with 1.32% egg white also had a significant effect on increasing the volume and decreasing the hardness of the rice bread (p<0.001). These results suggest that replacement of rice flour with 0.009% glucose oxidase and 0.006% fungal amylase, and 1.32% egg white are effective for producing rice bread with good volume and hardness.

The button mushroom, Agaricus bisporus, is one of the major economical crops cultivated in Korea. This mushroom showed the 5th production to 11,493 M/T in 2014. This study examined changes in decomposition-related enzyme activity secreted by the microorganisms isolated from button mushroom composts. Among the isolated microorganisms, bacterial strains CY-36 were the most dominant and had the highest enzyme activities. 16S rDNA sequencing analysis showed 99% similarity of CY-36 to Bacillus amyloliquefaciens. To define the optimum conditions for the mass production of CY-36, we have investigated optimum culture conditions and effects of various nutrient source on the bacterial growth. The optimum initial pH and temperature were determined as pH 5.0~8.0 and 30°C, respectively. The optimal culture medium for the growth was determined as follows: 2.0% dextrin, 1.0% yest extract, 0.5% NaNO3, 3 mM MgSO4, and 0.6% valine. Enzymatic activities of CMCase and PGase secreted by CY-36 were the most prominent and CMCase activity were optimal at 40°C and pH 7.0, and PGase activity were 50°C and pH 6.0. The activity of CMCase was rapidly decreased at more than 4°C and pH 5.0, and the activity of PGase was rapidly decreased at more than 30°C but didn’t affect as pH change. Accordingly, Bacillus amyloliquefaciens CY-36 was potentially useful on the fermentation stimulation and softening of rice straw in the process of fermentation for mushroom cultivation.

This study examined changes in decomposition-related enzyme activity secreted by the microorganisms isolated from button mushroom composts. Among the isolated microorganisms, bacterial strains CY-24 were the most dominant and had the highest enzyme activities. 16S rDNA sequencing analysis showed 97% similarity of CY-24 to Bacillus licheniformis. To define the optimum conditions for the mass production of CY-24, we have investigated optimum culture conditions and effects of various nutrient source on the bacterial growth. The optimum initial pH and temperature were determined as pH 6.0 and 30°C, respectively. The optimal culture medium for the growth was determined as follows: 0.3% mannose, 1.5% yest extract, 0.8% (NH4)2HPO4, 2 mM MnSO4, and 0.5% asparatic acid. Enzymatic activities of CMCase and PGase secreted by CY-24 were the most prominent and CMCase activity were optimal at 50~60°C and pH 6.0, and PGase activity were 60°C and pH 6.0. The activity of CMCase was inhibited by metal ions including Ca+ and Cu+, while the activity of PGase didn’t affect. The activity of CMCase was rapidly decreased at more than 30°C and pH 7.0, and the activity of PGase didn’t affect as temperature change but was rapidly decreased at more than pH 5.0. Accordingly, This strain was potentially useful on the fermentation stimulation and softening of rice straw in the process of fermentation for mushroom cultivation.

The hazards associated with the polycyclic aromatic hydrocarbons (PAHs) are known to be recalcitrant by their structure, but white rot fungi are capable of degrading recalcitrant organic compounds. Phlebia brevispora KUC9045 isolated from Korea was investigated its efficiency of degradation of four PAHs, such as phenanthrene, anthracne, fluoranthene, and pyrene. And the species secreted extracellular laccase and MnP (Manganese dependent peroxidase) during degradation. P. brevispora KUC9045 demonstrated effective degradation rates of phenanthrene (66.3%), anthracene (67.4%), fluoranthene (61.6%), and pyrene (63.3%), respectively. For enhancement of degradation rates of PAHs by the species, Remazol Brilliant Blue R (RBBR) was preferentially supplemented to induce ligninolytic enzymes. The biodegradation rates of the three PAHs including phenanthrene, fluoranthene, and pyrene were improved as higher concentration of Remazol Brilliant Blue R was supplemented. However, anthracene was degraded with the highest rate among four PAHs after two weeks of the incubation without RBBR addition. According to the previous study, RBBR can be clearly decolorized by P. brevispora KUC9045. Hence, the present study demonstrates simultaneous degradation of dye and PAHs by the white rot fungus. And it is considered that the ligninolytic enzymes are closely related with the degradation. In addition, it indicated that dye waste water might be used to induce ligninolytic enzymes for effective degradation of PAHs.

Basidiomycetous fungi are one of the most potent biodegraders because many of its species grow on dead wood or litter, in environments rich in lignocellulose. For the degradation of lignocellulose, basidiomycetes utilize their lignocellulytic enzymes, which typically include laccase (EC 1.10.3.2), lignin peroxidase (EC 1.11.1.14), xylanase (EC 3.2.1.8), and cellulase (EC 3.2.1.4). In recent years, the practical applications of basidiomycetes have ranged from the textile to the pulp and paper industries, and from food applications to bioremediation processes and industrial enzymatic saccharification of biomass. Recently, spent mushroom substrates of edible mushrooms have been used as sources of bulk enzymes to decolorize synthetic dyes in textile wastewater. In this review, the occurrence, mode of action, general properties, and production of lignocellulytic enzymes from mushroom species will be discussed. We will also discuss the potential applications of these enzymes

Proteolytic enzymes were screened to obtain anchovy protein hydrolysates (APH) with low-bitterness under pressurization. First, the residual proteolytic activity (%) and SDS-PAGE of commercially available food-grade protease (Alcalase® 2.4 L, FlavourzymeTM 500 MG, Neutrase®, ProtamexTM) and mixed enzyme (ME, Alcalase® : FlavourzymeTM = 1 : 5) after treatments of ambient pressure and 100 MPa at 50oC were investigated. The proteolytic activity of enzymes was still largely retained after pressure treatment. The SDS-PAGE on tested enzymes showed that the electrophoretic mobility maps (EMMs) after pressure treatment were similar to those at ambient pressure. With these results, it seemed that tested enzymes were stable at 100 MPa and 50oC. Raw anchovy hydrolyzed with various protease at 50oC under ambient pressure and 100 MPa for 1, 2, 6, 12, and 24 h. The degree of hydrolysis (DH, %) of prepared APHs at 100 MPa was increased, which suggests that the medium pressure treatment accelerates protein hydrolysis. The APH by ME exhibited the best suitable characteristics, which had a relatively higher DH (36.2±1.8- 57.5±2.8), a high content of oligopeptides with a molecular mass lower than 1,100 Da, mildly bitter taste as the bitterness value was lower than that of 0.2 g/L tea alkaloid, and the lowest contents of tryptophan. Thus, ME was selected to obtain APH with low-bitterness under pressurization.

This study was designed to investigate the effects of Korean Lycii fructus water extract in Al (Aluminum) administered rats. Forty-eight male Sprague-Dawley rats were divided into six groups: Control group, water extract group with 3% Lycii fructus, 1000 and 2000 ppm of Al groups, and 1000 and 2000 ppm of Al with 3% Lycii fructus water extract group. The Al content of rat tissue in the Al administered group was lower than that in rat tissue in the Al with 3% Lycii fructus water extract group. Plasma levels of renin and aldosterone activity was higher in the Al administration group, compared with the 3% Lycii fructus water extract group and Al administered group. Aspartate amino transaminase and alanine amino transaminase activities were elevated in the Al administered group and lower in the 3% Lycii fructus water extract group. Lactate dehydrogenase was lower in the 3% Lycii fructus water extract Al group than in the Al group. Choline acetyltransferase was higher in the 3% Lycii fructus water extract Al group than in the Al group.