Lipoxygenase(LOX) in soybeans is responsible for beany flavors which limit the wide utilization of soybeans to foods. This study was conducted to analyze beany flavor compounds of the normal Hwagkeumkong and LOX-deficient soybean cultivars, Jinpumkong which lacks L-2, L-3, and Jinpumkong 2 which lacks all L-1, L-2, L-3. Using the combination of dynamic headspace sampling and gas chromatography-mass selective detector(DHS-GC-MSD) for analyzing volatile compounds, hexanal and hexanol were identified in whole soy flour of all three soybena cultivars. Hwangkeumkong had more volatile compounds than Jinpumkong and Jinpumkong 2 in defatted soy flour. Hexanal and acetic acid were identified in soy milk of all three soybean cultivars but Hwangkeumkong had more volatile compounds than Jinpumkong 2. From the analysis with a static headspace sampling(SHS) and GC-MSD the major compounds were hexanal, acetic acid, 1-hexanol, and 1-octen-3-ol. The content of acetic acid was similar among three cultivars. But contents of hexanal and pentanal in Jinpumkong 2 were less than that of Jinpumkong and Hwangkeumkong. Using GC-FID, Jinpumkong 2 had less contents of hexanal and pentanol than Hwangkeumkong in whole soy flour and defatted soy flour. In this study, LOX-deficient soybean cultivars showed less hexanal, pentanol and other compounds than the normal Hwangkeumkong. However quite amount of beany flavor compounds were identified in Jinpumkong and Jinpumkong 2. So further studies are required to characterize LOX isozymes, to understand the mechanisms of beany flavors production, and to develop some other methods for removing beany flavor.
Lipoxygenase (LOX) is an industrial enzyme with wide applications in food and pharmaceutical industries. The available structure information indicates that eukaryotic LOXs consist of N terminus β-barrel and C terminus catalytic domains. However, the latest crystal structure of Pseudomonas aeruginosa LOX shows it is significantly different from those of eukaryotic LOXs, including the N-terminal helix domain. In this paper, the functions of this N-terminal helix domain in the soluble expression and catalysis of P. aeruginosa LOX were analyzed. Genetic truncation of this helix domain resulted in an insoluble P. aeruginosa LOX mutant. The active C-terminal domain was obtained by dispase digestion of the P. aeruginosa LOX derivative containing the genetically introduced dispase recognition sites. This functional C-terminal domain showed raised substrate affinity but reduced catalytic activity and thermostability. Crystal structure analyses demonstrate that the broken polar contacts connecting the two domains and the exposed hydrophobic substrate binding pocket may contribute to the insoluble expression of the C terminus domain and the changes in the enzyme properties. Our data suggest that the N terminus domain of P. aeruginosa LOX is required for its soluble expression in E. coli, which is different from that of the eukaryotic LOXs. Besides this, this N-terminal domain is not necessary for catalysis but shows positive effects on the enzyme properties. The results presented here provide new and valuable information on the functions of the N terminus helix domain of P. aeruginosa LOX and further improvement of its enzyme properties by molecular modification.
Changes of rancidity of soybean powder from Yeonhaeju (Bazaz) during storage were evaluated by TBA, and compared with those of Korean soybean (Baektae). Lipoxygenase (LOX) in the soybean powder was inactivated by high pressure processing (HPP) to increase the stability of soybean powder during storage. The level of inactivation of LOX was measured by spectrophotometer at 234 nm. HPP decreased the activity of LOX in the soybean powder of Baektae, compared to that of the control (i.e., soybean powder without HPP treatment) of Baektae, while TBA values of both HPP treated Baektae and the control were increased up to 24 days of storage. However, in case of Bazas, both LOX activity TBA values decreased after HPP treatment, compared to those of controls. The antioxidant compounds in both soybeans were measured and quantitatively evaluated by on-line $ABTS^{+}$ assay. Based on the trolox equivalent (TE) value at the retention time 38.2 and 40.1 min, the antioxidant components in Bazaz were higher than that of Baektae. It might indicate that relatively lower TBA values of HPP treated Bazaz was due to lower LOX activity as well as higher antioxidant compounds in the species.
Journal of the Korean Society of Food Science and Nutrition
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v.42
no.10
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pp.1629-1637
/
2013
The properties of non-GM soybeans with lipoxygenase (LOX) free genotypes, such as Gaechuck#1 (black, LOX2,3-free), Gaechuck#2 (yellow, LOX2,3-free) and Jinyangkong (yellow, LOX1,2,3-free), were compared to Taekwangkong (yellow, LOX1,2,3-present) as control. Length and weight of soybeans were significantly longer in Gaechuck#1, but LOX free genotypes of yellow beans was smaller than Taekwangkong, in terms the size and weight. After soaking for 12 hr, increased ratios of soybean weight were 217.07~226.55%, and they were significantly higher in Gaechuck#2. The total color difference (${\Delta}E$) to Taekwangkong was the highest in Gaechuck#1, but another yellow soybean were similar. Crude lipids content was significantly higher in Taekwangkong (17.30%), and these LOX free genotypes (13.71~14.66%) did not show significant difference. Crude protein contents were significantly higher in Gaechuck#1 (34.79%) compared to the other soybeans (32.93~33.47%). Contents of glutamic acid among the detected composition amino acid were highest, and its total contents were highest in Gaechuck#1 (32.42 g/100 g), and the total content of Gaechuck#2 was similar to Taekwangkong. Contents of total isoflavone were higher in LOX free genotypes than Taekwangkong, especially significantly higher in Gaechuck#1 (1672.54 ${\mu}g/g$). Contents of total phenol were highest in Gaechuck#1 (276.91 mg/100 g), and flavonoids content of Gaechuck#1 (26.93 mg/100 g) was 2.6~2.8 times higher than the other samples. DPPH, ABTS radical scavenging activities and reducing power were significantly higher in Gaechuck#1. The yield of soybean curd from tested samples was the highest in Gaechuck#2. Gaechuck#2 as LOX free genotypes was similar to Taekwangkong with regard to physicochemical properties and antioxidant activities. Therefore, we propose that Gaechuck#2 is a more suitable genotype for soybean curd products.
It is reported that the absence of lipoxygenase-3 (LOX-3) may contribute to a reduction in stale flavor after the storage of rice. To improve the quality of stored rice of the Korean japonica rice cultivar, we conducted a breeding program to develop near-isogenic rice without LOX-3 in the genetic background of Saenuri, a mega variety of Korea. In the first step of the breeding program, we used a donor parent of LOX-3 null, Daw Dam, and a recurrent japonica parent, Sindongjin, to develop HR27873-AC12 by backcross (BC1), color test for introgression of lox-3, and anther culture for rapid fixation. In the second step, we used the donor parent, HR27873-AC12, and the recurrent parent, Saenuri, to develop HR28896-31-3-1-1 by backcross (BC1), marker-assisted selection (MAS) for lox-3, and phenotypic selection (PS) for agronomic traits. Finally, in the third step, we developed HR30960-186-2-1-2-1 (Jeonju624), derived from a cross between Saenuri and HR28896-31-3-1-1, by MAS for lox-3 and PS with high selection pressure for agronomic characteristics. Jeonju624 was confirmed with the introgression of lox-3 by molecular marker. Jeonju624 was a mid-late maturing rice with similar agronomic characteristics to Saenuri, lodging tolerance with short culm, erect plant architecture, and resistance to bacterial blight and rice stripe virus. The yield components of Jeonju624 were mostly similar to Saenuri, except for the 1,000-grain weight of brown rice. The appearance of the grain of Jeonju624 was better than that of Saenuri, and the characteristics of cooked rice were similar to those of Saenuri. In the genetic background analysis using 406 KASP (Kompetitive Allele-Specific PCR) markers, Jeonju624 was confirmed to be the near-isogenic line (NIL) of Saenuri with a 95.8% recovery rate. Jeonju624 is the NIL of Saenuri without LOX-3, and overcomes the linkage drag of Daw Dam with similar agronomic characteristics and genetic background to Saenuri. Jeonju624 can be utilized as a practical cultivar to improve the quality of stored rice, breeding material for the introgression of lox-3, and genetic material to elucidate the effect of introgressed genes.
An investigation was conducted to ascertain the basic information on characteristics of embryonic tissue development among Korean Ginseng, American ginseng, $F_{1}$ hybrids and $BC_{1}F_{1}(F_{1}$ pollen back cross to $BC_{1}=Korean\;Ginseng)seeds$. The specific activities of lipoxygenase (LOX) and the protein assays were made during the above embryonic growth period at 5 or 25 degree C. The results are summarized as follows: The fresh weights(mg) of germinating seeds were significantly different among species. Ginseng seeds were stratified at $5^{\circ}C\;for\;60\;days$ the ratios of embryo/endosperms in $P.g.,\;P.q\;or\;F_{1}\;were\;about\;90{\%}$. The ratio was in $BC_{1}F_{1},\;79.2{\%}$ during the same period. The ratios of embryo/endosperms of seeds of P.g. or P.q. germinated at $5^{\circ}C$ showed rather higher values as compared to that at $25^{\circ}C$ LOX specific activity the highest in the $F_{1}$ and decreased in the order of P.g., P.q., and $BC_{1}F_{1}$. The highest LOX specific activity was shown at 80 days after sowing, followed by 70 days sowing, the least LOX activity was shown at 60 days.
Proceedings of the Korean Society of Crop Science Conference
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2017.06a
/
pp.69-69
/
2017
Due to the lack in storage facility, considerable amount of rice is stocked in the open air, which causes increased stale flavor and deteriorates palatability and merchantable quality. Lipoxygenase-3(LOX-3) is involved in the production of volatile constituents in stored rice, and the development of stale flavor is delayed in LOX-3 null rice. LOX activity in the rice grain is localized in bran fraction and the tropical Japonica cultivar 'Dawdam' was reported that the absence of LOX-3 is inherited as a simple recessive trait. Also, it was reported that the peroxidation of unsaturated fatty acids occurs at lower levels in the 'Dawdam' bran fraction during storage than in rice varieties with LOX-3. This study was conducted to develop LOX-3 null rice lines using 'Dawdam' and investigate changes of physicochemical properties of the lines stored at different storage temperatures and periods. So we analyzed texture, toyo glossiness value, germination rate and lipoxygenase activity of 15 LOX-3 null rice lines on the condition of which rough rice had been stored at different temperatures (high temperature condition at $35^{\circ}C$ and low temperature condition at $15^{\circ}C$ for 4months. Hardness and stickiness of the lines tendered to be increased when it was stored at high temperature and adhesiveness, springiness, cohesiveness and chewiness was not considerably different according to storage temperatures and periods. The germination rate of HR29062-B-98-2-1-B among LOX-3 null rice lines was higher than another lines, 99.3, 94.0% after 4months stored at low temperature and high temperature, respectively. The lipoxygenase activity was 3,304, 1,601unit/mg protein after 4months stored at low temperature and high temperatures, respectively. So, it is thought that this line will be useful to breed rice varieties with high storability after tested on agricultural traits.
Kim, In-Sung;Lee, Soo-Jung;Lee, Hye-Jin;Oh, Soo-Jeong;Chung, Jong-Il;Sung, Nak-Ju
Food Science and Preservation
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v.21
no.2
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pp.215-223
/
2014
The quality characteristics and antioxidant activity of three kinds of tofu made from lipoxygenase (LOX)-free genotypes soybeans were compared to the Taekwang (LOX-present) tofu as the control. The mineral contents of Jinyang tofu were significantly higher than those of the control. The hardness and gumminess of tofu made from LOX-free genotypes were significantly higher than those of the control. Gaechuck#2 tofu showed higher sensory evaluation results than the other kinds of tofu in terms of taste, flavor and overall acceptability. The total isoflavone contents were higher in the LOX-free genotypes than in the control. The total phenol content was similar for the Gaechuck#1, #2 and Taekwang tofu. The flavonoid content was higher in Gaechuck#1 and #2 tofu than in the control. The antioxidant activities were the highest in Gaechuck#1 tofu, followed by the Gaechuck#2. During the 15-day storage at $4^{\circ}C$, the turbidity of the immersing water tended to increase, but Gaechuck#1 and #2 tofu were significantly lower than in the control after 15-day storage. Therefore it is suggested that Gaechuck#2 tofu could be the suitable genotype for tofu products because it is the most effective in terms of overall acceptability, antioxidant activity, and storage stability.
The effect of lipoxygenase (LOX) on the oxidation and co-oxidation of lipid fraction was studied in the model system of rainbow trout. For the reaction in model system 1 g of lipid fraction and 50mL of enzyme extract(LOX, 140 unit in 50mL phosphate buffer solution at pH 7,4)), which were obtained from rainbow trout, were homoginized in the presence of Tween 20 and kept at 23$\circ$C for 3 days. The activity of LOX was decreased to 43% of initial level during the reaction in the model system. The initial composition of rainbow trout lipid was showed to be consisted of trigliceride(TG;82%) and free fatty acid(FFA;0.1%), while this converted to 59% of TG and 20% of FIFA, respectively after reaction in model system. Change of fatty acid composition was also observed and the content of linoleic acid, one of the major fatte acids, was decreased to 13% from 54% in the content of total fatty acids after reaction. The carotenoids in rainbow trout were composed of 0.4% $\alpha$-carotene, 1.6% $\beta$ -carotene, 80% canthaxanthin, 7% lutein and 11% zeaxanthin, thus the canthaxanthin was the major component. This canthaxanthin was the most degraded carotenoid by lipoxygenase catalyzed co-oxidation during the reaction. On the other hand the tocopherol isomers found in the rainbow trout were $\alpha$ and $\beta$ -tocopherol, and $\alpha$-tocopherol had a higher degradation rate by the lipoxygenase catalyzed co-oxidation than of $\beta$-tocopherol in the reaction of model system.
The role of intracellular $Ca^{2+}$, in the regulation of tumor cell proliferation by products of arachidonic acid (AA) metabolism was investigated using U-373 MG human as trocytoma cells. Treatment with nordihydroguaiaretic acid (NDGA), a lipoxygenase (LOX) inhibitor, or caffeic acid (CA), a specific 5-LOX inhibitor, suppressed proliferation of the tumor cells in a dose-dependent manner. However, indomethacin (indo), a cyclooxygenase (COX) inhibitor, did not significantly alter proliferation of the tumor cells. At anti-proliferative concentrations, NDGA and CA significantly inhibited intracellular $Ca^{2+}$ release induced by carbachol, a known intracelluar $Ca^{2+}$ agonist in the tumor cells. Exogenous administration of leukotriene $B_4(LTB_4)$, an AA metabolite of LOX pathway, enhanced proliferation of the tumor cells in a concentration-dependent fashion. In addition, $LTB_4$, induced intracelluar $Ca^{2+}$ release. Intracellular $Ca^{2+}$-inhibitors, such as an intracellular $Ca^{2+}$ chelator (BAPTA) and intracellular $Ca^{2+}$-release inhibitors (dantrolene and TMB-8), significantly blocked the LTB4-induced enhancement of cell proliferation and intracellular $Ca^{2+}$ release. These results suggest that LOX activity may be critical for cell proliferation of the human astrocytoma cells and that intracelluar $Ca^{2+}$ may play a major role in the mechanism of action of LOX.
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