Inhibitor cysteine knot (ICK) peptides exhibit ion channel blocking, insecticidal, and antimicrobial activities, but currently, no functional roles for bee-derived ICK peptides have been identified. In this study, a bee (Apis cerana) ICK peptide (AcICK) that acts as an antifungal peptide and as an insecticidal venom toxin was identified. AcICK contains an ICK fold that is expressed in the epidermis, fat body, or venom gland and is present as a 6.6-kDa peptide in bee venom. Recombinant AcICK peptide (expressed in baculovirus-infected insect cells) bound directly to Beauveria bassiana and Fusarium graminearum, but not to Escherichia coli or Bacillus thuringiensis. Consistent with these findings, AcICK showed antifungal activity, indicating that AcICK acts as an antifungal peptide. Furthermore, AcICK expression is induced in the fat body and epidermis after injection with B. bassiana. These results provide insight into the role of AcICK during the innate immune response following fungal infection. Additionally, we show that AcICK has insecticidal activity. Our results demonstrate a functional role for AcICK in bees: AcICK acts as an antifungal peptide in innate immune reactions in the body and as an insecticidal toxin in venom. The finding that the AcICK peptide functions with different mechanisms of action in the body and in venom highlights the two-pronged strategy that is possible with the bee ICK peptide.

Insect-derived Kazal-type serine protease inhibitors exhibit thrombin, elastase, plasmin, proteinase K, or subtilisin A inhibition activity, but so far, no functional roles for bee-derived Kazal-type serine protease inhibitors have been identified. In this study, a bee (Apis cerana) venom Kazal-type serine protease inhibitor (AcKTSPI) that acts as a microbial serine protease inhibitor was identified. AcKTSPI contained a single Kazal domain that displayed six conserved cysteine residues and a P1 threonine residue. AcKTSPI was expressed in the venom gland and was present as a 10-kDa peptide in bee venom. Recombinant AcKTSPI Kazal domain (AcKTSPI-Kd) expressed in baculovirus-infected insect cells demonstrated inhibitory activity against subtilisin A (Ki 67.03 nM) and proteinase K (Ki 91.53 nM), but not against α-chymotrypsin or typsin, which implies a role for AcKTSPI as a microbial serine protease inhibitor. However, AcKTSPI-Kd exhibited no detectable inhibitory effects on factor Xa, thrombin, tissue plasminogen activator, or elastase. Additionally, AcKTSPI-Kd bound directly to Bacillus subtilis, B. thuringiensis, Beauveria bassiana, and Fusarium graminearum but not to Escherichia coli. Consistent with these findings, AcKTSPI-Kd showed antibacterial activity against Gram-positive bacteria and antifungal activity against both plant-pathogenic and entomopathogenic fungi. These findings constitute molecular evidence that AcKTSPI acts as an inhibitor of microbial serine proteases. This paper provides a novel view of the antimicrobial functions of a bee venom Kazal-type serine protease inhibitor.

Worldwide studies on Apis cerana variation for biogeography and genetic diversity depended largely on a 86~93 bp-long mitochondrial non-coding region (internal spacer region) located between tRNALeu and COII (named as NC2), possibly due to higher variability among available markers. In order to incorporate the A. cerana occurring in South Korea into world extensive data, we also sequenced the NC2 from 118 A. cerana samples collected over nine Korean localities and 66 A. cerana samples over seven Asian localities, such as China, Vietnam, and Thailand. These data were combined with preexisting world data to scrutinize genetic relationships of A. cerana in South Korea to outside distributional range. Sequencing of 184 samples provided a total of ten haplotypes: five from Korea, six from China, one from Vietnam, and two from Thailand. Among them eight were new, whereas two were previously reported ones. Phylogenetic analysis of A. cerana NC2 haplotypes so far found including ours has confirmed the presence of four major groups of A. cerana (Asian mainland group, Sundaland group, Palawan group, and Luzon-Mindahnao group) and all haplotypes found in this study also were included in the Asian mainland group. In order to find further variable regions that can be used as sequence-based marker several mitochondrial non-coding regions and nuclear intron regions are in the middle of testing.

The bumblebee, Bombus ignitus (Hymenoptera: Apidae), is a valuable natural resource that is one of the most notably utilized for greenhouse pollination in Korea. In order to understand the nature of genetic relationships, gene flow, and population structure of the species we sequenced a partial COI gene of mitochondrial DNA (mtDNA) corresponding to “animal barcode” region and the complete internal transcribed spacer 2 (ITS2) of the nuclear ribosomal DNA (nrDNA) collected from Korean localities. Although the 658-bp long mtDNA sequence provided only six haplotypes with the maximum sequence divergence of 0.61% (4 bp), the ITS sequences provided 84 sequence types with the maximum sequence divergence of 1.02% (21 sites), confirming better applicability of the ITS sequences to the study of intraspecific variation. The complete ITS2 sequences of B. ignitus were shown to be longest among known insects, ranging in size from 2,034 bp ~ 2,052 bp, harboring two duplicated repeats. Overall, a very high per generation migration ratio, a very low level of genetic fixation, and no discernable hierarchical population/ population group were noted to exist among populations of B. ignitus on the basis of both molecules, thus suggesting that the B. ignitus populations on the Korean peninsula are panmictic, which is consistent with our understanding of the dispersal capability of the species

본 연구는 잠재적인 생균제제 Lactobacillus plantarum P1201에 의한 유색 소립콩 분말 두유의 젖산발효 중 이화학적 특성, 총 페놀릭스와 이소플라본 함량 및 항산화 활성(DPPH와 ABTS 라디칼 소거활성 및 FRAP 환원력) 변화를 연구하였다. 콩 분말 두유의 발효 중 pH는 감소하였으나, 산도, 생균수 및 β-glucosidase 활성은 증가하였다. 총 이소플라본 함량은 풍산나물콩이 다른 유색 소립콩보다 함량이 높았다. 발효과정에서 배당체 이소플라본인 daidzin과 genistin 함량은 감소하였고 이에 상응하여 비배당체 이소플라본인 daidzein과 genistein 함량은 급격히 증가하였다. 특히, 발효 60시간 후 발효된 풍산나물콩 분말 두유의 daidzein과 genistein 함량은 87.37 μg/g and 51.29 μg/g으로 다른 시료들보다 함량이 가장 높았다. 총 페놀릭스 함량은 다른이 다른 유색 소립콩보다 함량이 높았다. 총 페놀릭스 함량은 색 소립콩 분말 두유의 젖산발효 중 0.44-2.92 mg/g(0 h)에서 1.79-3.03 mg/g(60 h)로 급격히 증가하였다. 끝으로, 발효된 콩 분말 두유는 DPPH와 ABTS 라디칼 소거활성 및 FRAP 환원력은 각각 28.7-40.6에서 90-95.3%, 25.1-42.3에서 51.4-82.8% 및 0.73-1.54에서 0.98-1.79(OD593nm)로 증가하였다. 발효된 콩 분말 두유는 유색 소립콩의 가치를 증대시킬 수 있다.

This study aimed to produce fermented soy-powder milk (FSPM) with Lactobacillus plantarum P1201 and to evaluate its anti-obesity activity. Isoflavone and conjugated linoleic acid (CLA) of unfermented soy-powder milk (UFSPM) and FSPM and were analyzed via high-pressure liquid chromatography (HPLC) and gas chromatography (GC). Their inhibitory activities against α-glucosidase, α-amylase, and pancreatic lipase were assayed. Their anti-obesity activities were evaluated on the basis of their inhibitory effects on adipocyte differentiation in 3T3-L1 cells, and the expression of mRNAs associated with adipogenesis and lipid metabolism were analyzed via real time-polymerase chain reaction (RT-PCR) and quantitative PCR (qPCR). FSPM with L. plantarum P1201 increased the isoflavone aglycones (daidzein, glycitein, and genistein) content and produced CLA in soy-powder milk (SPM), both of which possessed bio-activity. Both UFSPM and FSPM showed dose-dependent inhibitory activity for α-glucosidase, α-amylase, and pancreatic lipase. FSPM, but not UFSPM, suppressed adipogenesis in 3T3-L1 cells and reduced their triglyceride content by 23.1% after treatment with 1,000 μg/mL of FSPM, compared with the control group. The anti-obesity effect of FSPM can be attributed to CLA and isoflavone aglycones, which targeted CCAAT/enhancer binding protein α (C/EBP-α) and down-regulated lipoprotein lipase (LPL), adiponectin, adipocyte fatty acid-binding protein (aP2), fatty acid synthase (FAS), and acetyl CoA carboxylase (ACC) mRNA. Furthermore, FSPM enhanced the inhibitory activity of glucosidase and pancreatic enzymes and anti-obesity activity. Further studies are required to investigate whether the anti-obesity effect of FSPM persists in an in vivo mouse model of diet-induced obesity.

This study investigated the nutritional properties and biological activities of Ganoderma lucidum (GL). The round type of GL contained higher carbohydrate content, while the Nokgak type of GL contained higher crude ash, crude fat, and crude protein content. The most abundant amino acid, fatty acid, mineral, and soluble vitamin observed were valine (round type: 11.90 mg/g and Nokgak type: 17.18 mg/g), linoleic acid (round type: 47.56% and Nokgak type: 75.68%), potassium (round type: 116.50 mg/100 g and Nokgak type: 184.36 mg/100 g), and vitamin B3 (round type: 1.78 mg/100 g and Nokgak type: 1.81 mg/100 g), respectively. In addition, the β-glucan content were 34.15 g/100 g (round type) and 30.07 g/100 g (Nokgak type). The GL 70% ethanol extract at 40℃ showed higher radical scavenging as well as carbohydrate and lipid enzyme inhibition than other conditions. At 1 mg/mL of treatment with the 70% ethanol extract at 40℃ of round type GL, the DPPH, ABTS, hydroxyl radical scavenging, and α-glucosidase, α-amylase, and pancreatic lipase inhibition activities obtained were approximately 92.85, 99.74, 58.09, 89.68, 44.68, and 67.56%, respectively.

In this study, vineger was produced after heat treatment of Elaeagnus multiflora juice and its fermentative characteristics were investigated. The heat-treated juice and vinegar of E. multiflora were similar in fruit color, with b values (redness) of 39.48 (juice) and 37.56 (vinegar). After 10 days of fermentation of E. multiflora fruit, the acetic acid bacteria viable cell number, pH, acidity, reducing sugar content, and alcohol content were 4.59-4.62 log CFU/mL, 3.14-3.45, 0.2-2.12%, 0.69-35.24 mg/mL, and 0.2%, respectively. The heat-treated juice and vinegar showed significantly higher radical scavenging and digestive enzyme inhibitory activities than untreated samples, and the levels of soluble phenolics, soluble flavonoids, flavan-3-ol derivatives, and phenolic and derivatives were increased. Additioinally, the heat-treated vinegar contained major organic acids, such as acetic acid (21.82 mg/mL), and major flavan-3-ols and phenolic acids, such as catechin (72.24 μg/mL), catechin gallate (273.36 μg/mL), epigallocatechin gallate (68.35 μg/mL), protocatechuic acid (12.84 μg/mL), and salicylic acid (42.29 μg/mL). At 25 μL/mL treatment, DPPH and ABTS radical scavenging activities and α-glucosidase and pancreatic lipase inhibitory activities were 79.66%, 93.99%, 90.12%, and 64.85%, respectively. This result suggested that it is possible to produce new types of vinegar and beverages, using heat-treated E. multiflora juice.

In this study, soy-powder yogurt (SPY) with enhanced levels of γ-aminobutyric acid (GABA) and isoflavone aglycone was produced from sprouting high-protein soybeans (HPSs). The fermented steam-HPS sprouts (0 to 4 cm) were fermented (72 h) with Lactobacillus brevis, and the total free amino acids (FAAs) of the formed mixtures were determined to be 79.53, 489.93, 877.55, 780.53, and 979.97 mg/100 mL in the fermented HPS (FHPS), and the fermented steam-HPS with 0 cm (FSHPS-0), 1 cm (FSHPS-1), 2 cm (FSHPS-2), and 4 cm sprouting lengths (FSHPS-4), respectively. The levels of glutamic acid (GA) and GABA were observed to be the highest, 100.31 and 101.60 mg/100 mL, respectively, in the unfermented HPS (UFSHPS-1, 1 cm) and FSHPS-1 sprouts, respectively. Moreover, the total contents of the isoflavone glycoside form decreased proportionally to the increasing total levels of isoflavone aglycones after fermentation in FSHPS-0, FSHPS-1, FSHPS-2, and FSHPS-4. The levels of isoflavone aglycones were detected as 350.34, 289.15, 361.61, 445.05, and 491.25 μg/g in FHPS, FSHPS-0, FSHPS-1, FSHPS-2, and FSHPS-4, respectively. While FSHPS-1 exhibited the highest DPPH (63.28%) and ABTS (73.28%) radical scavenging activities, FSHPS-4 contained the highest isoflavone aglycone ratio (81.63%). All in all, the FSHPS-1 mixture prepared in this study exhibited high GABA content and functional prosperity, thereby making it suitable for potential applications in the soy-dairy industry.

In this study, contetnts of phenolic acid and isoflavone, and biological activities of soy sauce were compared the soy sauce added bitter melon powder (BMPs). After the fermentation, pHs were decreased from 5.83 (0% BMP), 5.47 (5% BMP), and 5.32 (10% BMP) to 5.28, 5.36, and 5.16 at 90 days, whereas the acidities of soy sauce were increased from 0.06%, 0.07%, and 0.09% to 0.30%, 0.28%, and 0.36% at 90 days, respectively. In addition, the salinities of soy sauce were decreased, while viable cell numbers including Bacillus and yeast were increased. The contents of total phenolic, isoflavone-aglycone, and phenolic acid and antioxidant and α-glucosidase inhibition activities were significantly increased for 90 days, while the isoflavone-glycoside contents were decreased. In Particular, soy sauce with 10% BMP at 90 days showed the highest contents of glutamic acid (GA, 9,876.09 mg/100 mL) and γ-aminobutyric acid (GABA, 325.02 mg/100 mL) contents than among other samples. Additionally, the radical scavenging activities (DPPH, ABTS, ⦁OH, and FRAP) and α-glucosidase inhibition activities of soy sauce with 10% BMP at 90 days were shown to be high 96.07%, 97.27%, 59.47%, 1.98%, and 79.96%, respectively.

본 연구는 여주 분말 첨가 된장의 발효 중 이화학적 특성, phytochemical 화합물(isoflavones과 phenolic acids) 및 생리활성 활성 변화를 측정하였다. 된장 발효 중 pH는 6.41-5.83에서 5.81-5.24로 감소하였고, 반면에 산도는 0.42-0.65%에서 1.28-1.48%로 증가하였다. 고초균과 효모 생균수, 염도 및 총 아미노산 함량은 발효 종기(60 일)에 증가하였다. 특히, 발효 후 10% 여주 분말 첨가 된장은 가바(GABA, 129.87 mg/100 g) 함량이 다른 된장 시료들과 비교하여 가장 함량이 높았다. 발효 후 된장은 발효 전 된장 시료보다 생리활성이 높았다. 발효 후 phytochemicals 화합물 중 isoflavone-aglycones과 phenolic acids는 증가하였고, 반면에 isoflavone-glycosides는 감소하였다. 게다가, 발효 후에 총 phenolics, isoflavone-aglycones과 phenolic acids 함량은 급격히 증가하였고, 이에 따라 항산화 활성과 α-glucosidase 저해활성이 증가하였다. 이들 결과로부터 여주 분말 첨가로 항산화와 항당뇨 활성이 개선된 새로운 타입의 된장 제조가 가능할 것으로 판단되었다.

뜰보리수 열매 식초 발효 중 이화학적 특성, 생리활성물질(flavonols과 phenolic acids) 및 생리활성 변화를 조사하였다. 자연 발효 중 pH와 환원당은 발효 초기 각각 3.55와 6.88 mg/mL에서 3.34와 2.13 mg/mL로 감소하였고, 반면에 산도는 0.48%에서 5.48%로 증가하였다. 알코올 함량은 20일째까지 증가하여 6.6%로 최대치를 나타낸 후 감소하여 발효 종기 2.0%를 나타내었다. 초산균수는 발효 10일째 4.32 log CFU/mL에서 5.4 log CFU/mL로 증가하였고, 효모균수는 3.23 log CFU/mL에서 5.5 log CFU/mL로 증가하였다. 뜰보리수 열매 식초의 주요 유기산은 초산, 젖산 및 사과산으로 각각 38.84 mg/mL, 4.92 mg/mL 및 1.51 mg/mL를 함유하고 있었다. 수용성 phenolics와 flavonoids 함량은 발효 중에 증가하여 발효 초기 각각 0.79 mg/mL과 0.12 mg/mL에서 1.22 mg/mL과 0.14 mg/mL로 증가하였다. Flavonols 화합물 중 epicatechin gallate는 발효 10일째 168.1 μg/mL에서 115.97 μg/mL로 감소하였으나, phenolic acids 화합물 중 gallic acid는 18.52 μg/mL에서 95.07 μg/mL로 증가하였다. 자연 발효 60일째 DPPH 라디칼 소거활성은 71.35%, ABTS 라디칼 소거활성은 79.27%, hydroxyl 라디칼 소거활성은 68.72%, α-glucosidase 저해활성은 85.42%, α-amylase 저해활성은 52.12% 및 pancreatic lipase 저해활성은 53.66%를 나타내었다.