CAO Kun, WANG Ruonan, FANG Yali, et al. Investigation on the Effect of Temperature on the Activity of Lactate Dehydrogenase Based on Molecular Dynamics Simulation[J]. Science and Technology of Food Industry, 2022, 43(1): 134−140. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2021010025.
Citation: CAO Kun, WANG Ruonan, FANG Yali, et al. Investigation on the Effect of Temperature on the Activity of Lactate Dehydrogenase Based on Molecular Dynamics Simulation[J]. Science and Technology of Food Industry, 2022, 43(1): 134−140. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2021010025.

Investigation on the Effect of Temperature on the Activity of Lactate Dehydrogenase Based on Molecular Dynamics Simulation

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  • Received Date: January 08, 2021
  • Available Online: November 04, 2021
  • Lactate dehydrogenase (LDH) is one of the important enzymes for anaerobic glycolysis and gluconeogenesis. It could catalyze pyruvate to form lactic acid, which had high application value in food fermentation industry. However, LDH was easily affected by high temperature, which leaded to the decline of lactic acid production. To study the effects of different temperature on the conformation and activity of LDH, molecular dynamics simulations at four different temperatures (37, 55, 70 and 85 °C) were performed for 80 ns, respectively. The overall conformational changed and the differences of enzyme active centers were analyzed. The results showed that LDH was relatively stable at 37 and 55 °C, and the root mean square error, root mean square fluctuation, radius gyration and solvent accessible surface area of LDH increased significantly at 70 and 85 °C. Moreover, the secondary structure of protein had changed greatly at 85 °C, which indicated that high temperature would lead to protein conformational instability. Comparing the binding ability of pyruvate at 37 and 85 °C, which was the substrate of the enzyme, we found that high temperature would increase the distance between the residues of pyruvate binding sites, and then destroy the microenvironment of substrate molecular binding. Therefore, the denaturation of LDH occurred when the temperature exceeded 70 °C, and the denaturation degree was positively correlated with the increase of temperature. This would lead to the loss of its enzyme activity, which was not conducive to the application value in food industry. In this study, the effects of four different temperatures on LDH were analyzed at atomic level, and the key information of enzyme activity and conformation changes were revealed, which provided theoretical support for the selection of appropriate temperature in the fermentation process of lactic acid products.
  • [1]
    DELCENSERIE V, MARTEL D, LAMOUREUX M, et al. Immunomodulatory effects of probiotics in the intestinal tract[J]. Current Issues in Molecular Biology,2008,10(1-2):37−54.
    [2]
    ZHU Z Y, CUI D, GAO H, et al. Efficient synthesis and activity of beneficial intestinal flora of two lactulose-derived oligosaccharides[J]. European Journal of Medicinal Chemistry,2016,114:8−13. doi: 10.1016/j.ejmech.2016.03.007
    [3]
    WANG L, ZHANG Y, FAN G, et al. Effects of orange essential oil on intestinal microflora in mice[J]. Journal of the Science of Food and Agriculture,2019,99(8):4019−4028. doi: 10.1002/jsfa.9629
    [4]
    尤可言, 荒草. 自制酸奶和市售酸奶[J]. 少儿科技,2019,182,183(Z2):20−20. [YOU K Y, HUANG C. Homemade yogurt and commercially available yogurt[J]. Children's Science and Technology,2019,182,183(Z2):20−20.
    [5]
    刘学云, 于新, 何嘉敏, 等. 九种益生菌之间的相互作用及协同共生机理[J]. 食品与发酵工业,2019,45(13):65−70. [LIU X Y, YU X, HE J M, et al. Interactions between nine probiotics and mechanisms of cooperative symbiosis[J]. Food and Fermentation Industry,2019,45(13):65−70.
    [6]
    王希, 洪鲲, 赵玉丹, 等. 乳酸发酵第2阶段能量释放生物学教学研究[J]. 生物学通报,2019(2):43−45. [WANG X, HONG K, ZHAO Y D, et al. Teaching research on energy release biology in the second stage of lactic acid fermentation[J]. Biology Bulletin,2019(2):43−45. doi: 10.3969/j.issn.0006-3193.2019.02.018
    [7]
    BUJNA E, NIKOLETTA A F, TRAN A M, et al. Lactic acid fermentation of apricot juice by mono- and mixed cultures of probiotic Lactobacillus and Bifidobacterium strains[J]. Food Science and Biotechnology,2018,27(2):547−554.
    [8]
    GODERSKA K. The antioxidant and prebiotic properties of lactobionic acid[J]. Applied Microbiology & Biotechnology,2019,103(9):3737−3751.
    [9]
    WANGA M, CHENA Y, WANGC Y, et al. Beneficial changes of gut microbiota and metabolism in weaned rats with Lactobacillus acidophilus NCFM and Bifidobacterium lactis Bi-07 supplementation[J]. Journal of Functional Foods,2018,48:252−265. doi: 10.1016/j.jff.2018.07.008
    [10]
    钟秀斌, 邓申彪, 沈洋. 一种乳酸菌发酵的温度控制装置: 中国, CN211199202U[P], 2020.08.

    ZHONG X B, DENG S B, SHEN Y. A temperature control device for lactic acid bacteria fermentation: China, CN21119902u [P]. 2020.08.
    [11]
    岳林芳, 王俊国, 萨如拉, 等. 培养条件对乳酸菌发酵剂抗冷冻干燥性能影响的研究进展[J]. 食品科学,2016,7(11):270−276. [YUE L F, WANG J G, SA R L, et al. Effects of culture time, temperature and medium composition on freeze drying resistance of lactic acid bacteria starter[J]. Food Science,2016,7(11):270−276. doi: 10.7506/spkx1002-6630-201611047
    [12]
    侯若冰, 陈志达, 卞江, 等. L-乳酸脱氢酶催化反应机理的理论研究进展[J]. 化学通报,2000(1):15−21. [HOU R B, CHEN Z D, BIAN J, et al. Progress in theoretical research on catalytic reaction mechanism of L-lactate dehydrogenase[J]. Chemical Bulletin,2000(1):15−21. doi: 10.3969/j.issn.0441-3776.2000.01.004
    [13]
    DE BEER D, TOBIN J, WALCZAK B, et al. Phenolic composition of rooibos changes during simulated fermentation: Effect of endogenous enzymes and fermentation temperature on reaction kinetics[J]. Food Research International,2019,121(JUL.):185−196.
    [14]
    蔡沛蓉, 冯楠楠, 郑豪, 等. 玉米赤霉烯酮对大鼠睾丸支持细胞乳酸产生及相关蛋白表达的影响[J]. 南京农业大学学报,2019,42(5):911−916. [CAI P R, FENG N N, ZHENG H, et al. Effect of zearalenone on lactic acid production and expression of related proteins in Sertoli cells[J]. Journal of Nanjing Agricultural University,2019,42(5):911−916. doi: 10.7685/jnau.201812027
    [15]
    鲍志伟, 苏晓, 杨柳婷, 等. 重组大肠杆菌全细胞合成D-苯基乳酸[J]. 食品与发酵工业,2019,45(1):49−53. [BAO Z W, SU X, YANG L T, et al. Biocatalytic production of D-phenyllactic acid by using whole cells of recombinant Escherichia coli[J]. Food and Fermentation Industries,2019,45(1):49−53.
    [16]
    BLECKWEDEL J, MOHAMED F, MOZZI F, et al. Major role of lactate dehydrogenase D-LDH1 for the synthesis of lactic acid in Fructobacillus tropaeoli CRL 2034[J]. Applied Microbiology and Biotechnology,2020,104(3):7409−7426.
    [17]
    ANDRES J, MOLINER V, KRECHL J, et al. A PM3 quantum chemical study of the pyruvate reduction mechanism catalyzed by lactate dehydrogenase[J]. Bioorganic Chemistry,1993,21(3):260−274. doi: 10.1006/bioo.1993.1022
    [18]
    ASLAN A S, BIRMINGHAM W R, KARAGULER N G, et al. Semi-rational design of Geobacillus stearothermophilus l-lactate dehydrogenase to access various chiral alpha-Hydroxy acids[J]. Applied Biochemistry and Biotechnology,2016,179:474−484. doi: 10.1007/s12010-016-2007-x
    [19]
    ZHANG Y, TAN H, ZHAO J X, et al. Structural change from homogenous structure to staging in benzoic acid intercalated LDH: Experimental and molecular dynamics simulation insights[J]. Physical Chemistry Chemical Physics,2012,14(25):9067. doi: 10.1039/c2cp40674h
    [20]
    CAO K, LI N, WANG H, et al. Two zinc-binding domains in the transporter AdcA from Streptococcus pyogenes facilitate high-affinity binding and fast transport of zinc[J]. Journal of Biological Chemistry,2018:6075−6089.
    [21]
    CAO K, ZHANG J, MIAO X Y, et al. Evolution and molecular mechanism of PitAs in iron transport ofStreptococcus species[J]. Journal of Inorganic Biochemistry,2018:113−123.
    [22]
    曹剑, 曹赞霞, 赵立岭, 等. 分子动力学模拟Cu2+α-突触核蛋白(1-17)肽段构象变化的影响[J]. 物理化学学报,2012(2):479−488. [CAO J, CAO Z X, ZHAO L L, et al. Effect of Cu2+ on conformational changes of α-synuclein (1-17) peptide by molecular dynamics simulation[J]. Acta Physicochemical Sinica,2012(2):479−488. doi: 10.3866/PKU.WHXB201111231
    [23]
    VERMAAS J V, HARDY D J, STONE J E, et al. TopoGromacs: Automated topology conversion from CHARMM to Gromacs within VMD[J]. Journal of Chemical Information and Modeling,2016,56(6):1112−1116. doi: 10.1021/acs.jcim.6b00103
    [24]
    ADAMS M J, FORD G C, KOEKOEK R, et al. Structure of lactate dehydrogenase at 2.8 Å resolution[J]. Nature,1972,227(5263):1098−1103.
    [25]
    ZHENG Y, GUO S, GUO Z, et al. Effects of N-terminal deletion mutation on rabbit muscle lactate dehydrogenase[J]. Biochemistry (00062979),2004,69(4):401−406.
    [26]
    UCHIKOBA H, FUSHINOBU S, WAKAGI T, et al. Crystal structure of non-allosteric L-lactate dehydrogenase from Lactobacillus pentosus at 2.3 A resolution: specific interactions at subunit interfaces[J]. Proteins-structure Function & Bioinformatics,2010,46(2):206−214.
    [27]
    ARAI K, ISHIMITSU T, FUSHINOBU S, et al. Active and inactive state structures of unliganded Lactobacillus casei allosteric L-lactate dehydrogenase[J]. Proteins-structure Function & Bioinformatics,2010,78(3):681−694.
    [28]
    冯涛, 刘芳芳, 荣志伟, 等. 基于分子动力学模拟的直链淀粉风味分子包合物形成机理的研究[J]. 现代食品科技,2015(3):126−132. [FENG T, LIU F F, RONG Z W, et al. Formation mechanism of amylose flavor molecular inclusion complex based on molecular dynamics simulation[J]. Modern Food Science and Technology,2015(3):126−132.
    [29]
    GREEN S R, STOREY K B. Regulation of crayfish, Orconectes virilis, tail muscle lactate dehydrogenase (LDH) in response to anoxic conditions is associated with alterations in phosphorylation patterns[J]. Comparative Biochemistry & Physiology Part B,2016,202:67−74.
    [30]
    陈娇, 王玉丽, 徐为人, 等. 分子动力学模拟法研究糖类衍生物与钠-葡萄糖协同转运蛋白2的相互作用[J]. 中草药,2013,44(10):1440−1447. [CHEN J, WANG Y L, XU W R, et al. Study on the interaction between carbohydrate derivatives and sodium glucose cotransporter 2 by molecular dynamics simulation[J]. Chinese Herbal Medicine,2013,44(10):1440−1447.
    [31]
    丁伟, 刘国宇, 于涛, 等. 分子动力学模拟HEWL晶体在不同环境中的动力学行为[J]. 计算机与应用化学,2010,27(2):173−178. [DING W, LIU G Y, YU T, et al. The molecular dynamics simulation of HEWL in different conditions[J]. Computers and Applied Chemistry,2010,27(2):173−178. doi: 10.3969/j.issn.1001-4160.2010.02.009
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