植物中松脂醇-落叶松脂素还原酶催化特征研究进展

doi:10.11983/CBB22160

植物学报 ›› 2023, Vol. 58 ›› Issue (4): 656-667.DOI: 10.11983/CBB22160

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植物中松脂醇-落叶松脂素还原酶催化特征研究进展

阴艳红¹, 陈万生¹^,²(), 肖莹¹()

1.上海中医药大学中药研究所, 中药新资源与品质评价中管局重点研究室, 中药资源与生物技术中心, 上海 201203
2.海军军医大学长征医院药学部, 上海 200003

收稿日期:2022-07-20 接受日期:2022-12-13 出版日期:2023-07-01 发布日期:2023-01-10
通讯作者: *E-mail: chenwansheng@shutcm.edu.cn;xiaoyingtcm@shutcm.edu.cn
基金资助:
国家自然科学基金(81874335);国家自然科学基金(32170402);国家自然科学基金(31872665);2022重点研发计划中药现代化专项(2022YFC3501700);上海市2023年度科技创新行动计划青年优秀学术带头人项目(23XD1423500);上海市中央引导地方科技发展资金(YDZX20203100002948)

Research Progress on Catalytic Characteristics of Pinoresinol-lariciresinol Reductase in Plants

Yanhong Yin¹, Wansheng Chen¹^,²(), Ying Xiao¹()

1. Research and Development Center of Chinese Medicine Resources and Biotechnology, The SATCM Key Laboratory for New Resources & Quality Evaluation of Chinese Medicine, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
2. Department of Pharmacy, Changzheng Hospital, Naval Medical University, Shanghai 200003, China

Received:2022-07-20 Accepted:2022-12-13 Online:2023-07-01 Published:2023-01-10
Contact: *E-mail: chenwansheng@shutcm.edu.cn;xiaoyingtcm@shutcm.edu.cn

摘要/Abstract

摘要： 松脂醇-落叶松脂素还原酶(PLR)是植物中木脂素生物合成的关键酶, 能够催化松脂醇转化为落叶松脂素, 并进一步催化落叶松脂素生成开环异落叶松脂素, 且存在底物立体选择性, 是一种NADPH依赖型还原酶。PLR的催化产物位于不同类型8-8′木脂素的源头, 其底物选择性直接决定木脂素的骨架类型, 如呋喃、二苄基丁烷、二苄基丁内酯和芳基四氢萘木脂素。因此, PLR的催化特性和表达特征在植物木脂素组成及其生物活性多样性中发挥重要作用。该文综述了PLR在植物木脂素生物合成中的作用、对映异构体选择性及其催化机制, 以期为进一步研究PLR基因的生物学功能以及催化机制奠定基础, 并为不同类型木脂素的精确生物合成指明方向。

关键词: 松脂醇-落叶松脂素还原酶, 木脂素, 催化特征, 对映体选择性, 催化机制

Abstract: Pinoresinol-lariciresinol reductases (PLRs) are key enzymes involved in the lignan biosynthesis in plants, which convert pinoresinol to lariciresinol and then to secoisolariciresinol. PLRs are NADPH-dependent reductases with substrate stereoselectivity. The catalytic products of PLR are the sources of different types of 8-8′ lignans, and the substrate selectivity directly determines the skeleton types of lignans, such as furano, dibenzylbutane, dibenzylbutyrolactone and aryltetrahydronaphthalene lignans. Therefore, the catalytic and expression characteristics of PLRs play an important role in the composition and biodiversity of lignans in plants. This paper reviewed the research progress on the important role of PLRs in lignans biosynthesis, as well as its enantioselectivity and catalytic mechanism, thus to lay the foundation for further study on the biological function and catalytic mechanism of PLR genes, and point out the direction for the precise biosynthesis of different types of lignans enantiomers through synthetic biology.

Key words: pinoresinol-lariciresinol reductases (PLRs), lignans, catalytic characteristics, enantioselectivity, catalytic mechanism

阴艳红, 陈万生, 肖莹. 植物中松脂醇-落叶松脂素还原酶催化特征研究进展. 植物学报, 2023, 58(4): 656-667.

Yanhong Yin, Wansheng Chen, Ying Xiao. Research Progress on Catalytic Characteristics of Pinoresinol-lariciresinol Reductase in Plants. Chinese Bulletin of Botany, 2023, 58(4): 656-667.

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图/表 5

图1 木脂素生物合成途径 PAL: 苯丙氨酸解氨酶; C4H: 肉桂酸-4-羟化酶; 4CL: 4-香豆酸:辅酶A连接酶; HCT: 羟基肉桂酰转移酶; C3H: 香豆酸-3-羟化酶; CCoAOMT: 咖啡酰辅酶A-O-甲基转移酶; CCR: 肉桂酰辅酶A还原酶; CAD: 肉桂醇脱氢酶; DIR: 聚合蛋白酶; PLR: 松脂醇-落叶松脂素还原酶; SDH: 开环异落叶松脂醇脱氢酶; UGT: 依赖尿苷二磷酸的糖基转移酶

Figure 1 Lignan biosynthetic pathways PAL: Phenylalanine ammonia-lyase; C4H: Cinnamate-4-hydroxylase; 4CL: 4-coumarate:coenzyme A ligase; HCT: Hydroxycinnamoyltransferase; C3H: Coumarate-3-hydroxylase; CCoAOMT: Caffeoyl coenzyme A-O-methyltransferase; CCR: Cinnamoyl-CoA reductase; CAD: Cinnamyl alcohol dehydrogenase; DIR: Dirigent protein; PLR: Pinoresinol-lariciresinol reductase; SDH: Secoisolariciresinol dehydrogenase; UGT: Uridine diphosphatedependent glycosyltransferase

表1 植物中的PLR基因

Table 1 PLR genes from different plant species

植物名称	基因名称	GenBank 登录号	cDNA长度(bp)	开放阅读框长度(bp)	参考文献
金钟连翘(Forsythia intermedia)	FiPLR1	U81158	1047	939	Dinkova-Kostova et al., 1996
拟南芥(Arabidopsis thaliana)	AtPrR1	NM102944	1129	954	Nakatsubo et al., 2008
拟南芥(Arabidopsis thaliana)	AtPrR2	NM117440	1206	954	Nakatsubo et al., 2008
菘蓝(Isatis indigotica)	IiPLR1	JF264893	1062	954	Xiao et al., 2015
亚麻(Linum usitatissimum)	LuPLR1	AJ849359	-	939	von Heimendahl et al., 2005
亚麻(Linum usitatissimum)	LuPLR2	EU029951	1203	993	Hemmati et al., 2010
白亚麻(L. album)	LaPLR1	AJ849358	1482	981	von Heimendahl et al., 2005
黄亚麻(L. flavum)	LfPLR	MK599138	-	975	Akira et al., 2016
长萼亚麻(L. corymbulosum)	LcPLR1	EU107358	1244	948	Bayindir et al., 2010
宿根亚麻(L. perenne)	LpPLR1	EF050530	1145	945	Hemmati et al., 2007
桃儿七(Sinopodophyllum hexandrum)	ShPLR	EU855792	983	936	Wankhede et al., 2013
六角莲(Dysosma pleiantha)	DpPLR	KJ000045	-	933	Kuo et al., 2014
北美乔柏(Thuja plicata)	TpPLR1	AF242503	1190	942	Fujita et al., 1999
	TpPLR2	AF242504	1151	939
	TpPLR3	AF242505	1308	945
	TpPLR4	AF242506	1287	939
台湾杉(Taiwania cryptomerioides)	TcPLR1	MG264424	-	975	Chiang et al., 2018
	TcPLR2.2	MG264425	1138	942
	TcPLR3	MG264426	1102	939
山茶(Camellia sinensis)	CsPLR1	MH037247	1325	939	Wu et al., 2019
山茶(Camellia sinensis)	CsPLR2	MH037248	1126	939	Wu et al., 2019

图2 PLRs蛋白氨基酸序列及二级结构域比对(GXXGXXG: NADPH结合基序)

Figure 2 Comparison of amino acid sequences and secondary structure domain of PLRs (GXXGXXG: NADPH-binding motif)

图3 植物中的PLRs系统发育树及其对映体选择性 PIN: 松脂醇; LAR: 落叶松脂素; SEC: 开环异落叶松脂素。右侧为植物中的PLR对松脂醇和落叶松脂素对映体选择性的具体表现形式(/表示无催化活性)

Figure 3 Phylogenetic tree and enantioselectivity of PLRs in plants PIN: Pinoresinol; LAR: Lariciresinol; SEC: Secoisolariciresinol. On the right side, specific forms of enantioselectivity of PLRs in plants to pinoresinol and lariciresinol ( / indicate no catalytic activity)

图4 IiPLR1_NAP_+PIN蛋白晶体结构(A)和底物结合凹槽细节(B) α: α-螺旋; β: β-折叠。NADPH和+PIN的棒状结构分别为橙色和黄色; N-端的NADP+结合域(NBD)和C-端的底物结合域(SBD)分别为淡绿色和姜黄色; 虚线为+PIN与其周围活性氨基酸可能存在的氢键作用; Mol-A和Mol-B为同源二聚体的2个单体。

Figure 4 Protein crystal structure of IiPLR1_NAP_+PIN (A) and detail of substrate bonding groove (B) α: α-helix; β: β-sheet. NADPH and +PIN were highlighted in orange and yellow, respectively; the N-terminal NADP+ binding domain (NBD) and the C-terminal substrate binding domain (SBD) were colored in light green and ginger, respectively; dotted lines denote possible hydrogen bonds between +PIN and active residues around substrate; Mol-A and Mol-B are two monomer of homodimer.

参考文献 53

[1]	陈瑞兵 (2018). Dirigent蛋白催化菘蓝有效成分木脂素生物合成的机制研究. 博士论文. 上海: 中国人民解放军海军军医大学. pp. 20-152.
[2]	程丽姣, 丁羽佳, 翟永功, 赵长琦 (2006). 植物中新的木脂素类化合物及其生物活性. 国外医药·植物药分册 21(3), 93-100.
[3]	刘长军, 侯嵩生 (1997). 抗癌活性物质鬼臼类木脂素的研究进展. 天然产物研究与开发 9(3), 81-89.
[4]	Allen KL, Tschantz DR, Awad KS, Lynch WP, Delucia AL (2007). A plant lignan, 3′-O-methyl-nordihydroguaiaretic acid, suppresses papillomavirus E6 protein function, stabilizes p53 protein, and induces apoptosis in cervical tumor cells. Mol Carcinog 46, 564-575. DOI URL
[5]	Ayella AK, Trick HN, Wang WQ (2007). Enhancing lignan biosynthesis by over-expressing pinoresinol lariciresinol reductase in transgenic wheat. Mol Nutr Food Res 51, 1518-1526. PMID
[6]	Bayindir Ü, Alfermann AW, Fuss E (2008). Hinokinin biosynthesis in Linum corymbulosum Reichenb. Plant J 55, 810-820. DOI URL
[7]	Bohlin L, Rosen B (1996). Podophyllotoxin derivatives: drug discovery and development. Drug Discovery Today 1, 343-351. DOI URL
[8]	Calado A, Neves PM, Santos T, Ravasco P (2018). The effect of flaxseed in breast cancer: a literature review. Front Nutr 5, 4. DOI PMID
[9]	Céspedes CL, Avila JG, García AM, Becerra J, Flores C, Aqueveque P, Bittner M, Hoeneisen M, Martinez M, Silva M (2006). Antifungal and antibacterial activities of Araucaria araucana (Mol.) K. Koch heartwood lignans. Z Naturforsch C J Biosci 61, 35-43. DOI PMID
[10]	Chen X, Chen JF, Feng JX, Wang Y, Li SN, Xiao Y, Diao Y, Zhang L, Chen WS (2021). Tandem UGT71B5s catalyze lignan glycosylation in Isatis indigotica with substrates promiscuity. Front Plant Sci 12, 637695. DOI URL
[11]	Chiang NT, Ma LT, Lee YR, Tsao NW, Yang CK, Wang SY, Chu FH (2018). The gene expression and enzymatic activity of pinoresinol-lariciresinol reductase during wood formation in Taiwania cryptomerioides Hayata. Holzforschung 73, 197-208. DOI URL
[12]	Corbin C, Drouet S, Markulin L, Auguin D, Lainé É, Davin LB, Cort JR, Lewis NG, Hano C (2018). A genome-wide analysis of the flax (Linum usitatissimum L.) dirigent protein family: from gene identification and evolution to differential regulation. Plant Mol Biol 97, 73-101. DOI
[13]	Corbin C, Drouet S, Mateljak I, Markulin L, Decourtil C, Renouard S, Lopez T, Doussot J, Lamblin F, Auguin D, Lainé E, Fuss E, Hano C (2017). Functional characterization of the pinoresinol-lariciresinol reductase-2 gene reveals its roles in yatein biosynthesis and flax defense response. Planta 246, 405-420. DOI PMID
[14]	Cullmann F, Becker H (1999). Lignans from the liverwort Lepicolea ochroleuca. Phytochemistry 52, 1651-1656. DOI URL
[15]	Davin LB, Lewis NG (1992). Phenylpropanoid metabolism:biosynthesis of monolignols, lignans and neolignans, lignins and suberins. In: Stafford HA, Ibrahim RK, eds. Phenolic Metabolism in Plants. Boston: Springer. pp. 325-375.
[16]	Dinkova-Kostova AT, Gang DR, Davin LB, Bedgar DL, Chu A, Lewis NG (1996). (+)-pinoresinol/(+)-lariciresinol reductase from Forsythia intermedia. Protein purification, cDNA cloning, heterologous expression and comparison to isoflavone reductase. J Biol Chem 271, 29473-29482. DOI PMID
[17]	Favela-Hernández JMJ, García A, Garza-González E, Rivas-Galindo VM, Camacho-Corona MR (2012). Antibacterial and antimycobacterial lignans and flavonoids from Larrea tridentata. Phytother Res 26, 1957-1960. DOI PMID
[18]	Fujita M, Gang DR, Davin LB, Lewis NG (1999). Recombinant pinoresinol-lariciresinol reductases from western red cedar (Thuja plicata) catalyze opposite enantiospecific conversions. J Biol Chem 274, 618-627. DOI PMID
[19]	Gordaliza M, García PA, del Corral JMM, Castro MA, Gómez-Zurita MA (2004). Podophyllotoxin: distribution, sources, applications and new cytotoxic derivatives. Toxicon 44, 441-459. DOI PMID
[20]	Guo R, Lv TM, Shang XY, Yao GD, Lin B, Wang XB, Huang XX, Song SJ (2019). Racemic neolignans from Crataegus pinnatifida: chiral resolution, configurational assignment, and cytotoxic activities against human hepatoma cells. Fitoterapia 137, 104287. DOI URL
[21]	Hano C, Corbin C, Drouet S, Quéro A, Rombaut N, Savoire R, Molinié R, Thomasset B, Mesnard F, Lainé E (2017). The lignan (+)-secoisolariciresinol extracted from flax hulls is an effective protectant of linseed oil and its emulsion against oxidative damage. Eur J Lipid Sci Technol 119, 1600219. DOI URL
[22]	Hano C, Martin I, Fliniaux O, Legrand B, Gutierrez L, Arroo RRJ, Mesnard F, Lamblin F, Lainé E (2006). Pinoresinol-lariciresinol reductase gene expression and secoisolariciresinol diglucoside accumulation in developing flax (Linum usitatissimum) seeds. Planta 224, 1291-1301. DOI PMID
[23]	Hemmati S, Schmidt TJ, Fuss E (2007). (+)-pinoresinol/ (-)-lariciresinol reductase from Linum perenne Himmelszelt involved in the biosynthesis of justicidin B. FEBS Lett 581, 603-610. PMID
[24]	Hemmati S, von Heimendahl CB, Klaes M, Alfermann AW, Schmidt TJ, Fuss E (2010). Pinoresinol-lariciresinol reductases with opposite enantiospecificity determine the enantiomeric composition of lignans in the different organs of Linum usitatissimum L. Planta Med 76, 928-934. DOI PMID
[25]	Kassuya CAL, Leite DFP, de Melo LV, Rehder VLG, Calixto JB (2005). Anti-inflammatory properties of extracts, fractions and lignans isolated from Phyllanthus amarus. Planta Med 71, 721-726. DOI URL
[26]	Kraus C, Spiteller G (1997). Comparison of phenolic compounds from galls and shoots of Picea glauca. Phytochemistry 44, 59-67. DOI URL
[27]	Kumar S, Stecher G, Tamura K (2016). MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33, 1870-1874. DOI PMID
[28]	Kuo HJ, Wei ZY, Lu PC, Huang PL, Lee KT (2014). Bioconversion of pinoresinol into matairesinol by use of recombinant Escherichia coli. Appl Environ Microbiol 80, 2687-2692. DOI URL
[29]	Kuo YC, Kuo YH, Lin YL, Tsai WJ (2006). Yatein from Chamaecyparis obtusa suppresses herpes simplex virus type 1 replication in HeLa cells by interruption the immediate-early gene expression. Antiviral Res 70, 112-120. DOI URL
[30]	Lau W, Sattely ES (2015). Six enzymes from mayapple that complete the biosynthetic pathway to the etoposide aglycone. Science 349, 1224-1228. DOI PMID
[31]	Li KM, Dong X, Ma YN, Wu ZH, Yan YM, Cheng YX (2019). Antifungal coumarins and lignans from Artemisia annua. Fitoterapia 134, 323-328. DOI URL
[32]	Min T, Kasahara H, Bedgar DL, Youn B, Lawrence PK, Gang DR, Halls SC, Park H, Hilsenbeck JL, Davin LB, Lewis NG, Kang C (2003). Crystal structures of pinoresinol-lariciresinol and phenylcoumaran benzylic ether reductases and their relationship to isoflavone reductases. J Biol Chem 278, 50714-50723. DOI PMID
[33]	Nakatsubo T, Mizutani M, Suzuki S, Hattori T, Umezawa T (2008). Characterization of Arabidopsis thaliana pinoresinol reductase, a new type of enzyme involved in lignan biosynthesis. J Biol Chem 283, 15550-15557. DOI PMID
[34]	Rahman MMA, Dewick PM, Jackson DE, Lucas JA (1990). Lignans of Forsythia intermedia. Phytochemistry 29, 1971-1980. DOI URL
[35]	Renouard S, Tribalatc MA, Lamblin F, Mongelard G, Fliniaux O, Corbin C, Marosevic D, Pilard S, Demailly H, Gutierrez L, Hano C, Mesnard F, Lainé E (2014). RNAi-mediated pinoresinol lariciresinol reductase gene silencing in flax (Linum usitatissimum L.) seed coat: consequences on lignans and neolignans accumulation. J Plant Physiol 171, 1372-1377. DOI URL
[36]	Robert X, Gouet P (2014). Deciphering key features in protein structures with the new ENDscript server. Nucleic Acids Res 42, W320-W324. DOI URL
[37]	Scher JM, Zapp J, Becker H (2003). Lignan derivatives from the liverwort Bazzania trilobata. Phytochemistry 62, 769-777. DOI URL
[38]	Shiaishi A, Murata J, Matsumoto E, Matsubara S, Ono E, Satake H (2016). De novo transcriptomes of Forsythia koreana using a novel assembly method: insight into tissue- and species-specific expression of lignan biosynthesis-related gene. PLoS One 11, e0164805.
[39]	Stadler D, Bach T (2008). Concise stereoselective synthesis of (-)-podophyllotoxin by an intermolecular iron(III)- catalyzed Friedel-Crafts alkylation. Angew Chem Int Ed Engl 47, 7557-7559. DOI URL
[40]	Suzuki S, Umezawa T, Shimada M (2002). Stereochemical diversity in lignan biosynthesis of Arctium lappa L. Biosci Biotechnol Biochem 66, 1262-1269. DOI URL
[41]	Takeda R, Hasegawa J, Shinozaki M (1990). The first isolation of lignans, megacerotonic acid and anthocerotonic acid, from non-vascular plants, anthocerotae (hornworts). Tetrahedron Lett 31, 4159-4162. DOI URL
[42]	Teponno RB, Kusari S, Spiteller M (2016). Recent advances in research on lignans and neolignans. Nat Prod Rep 33, 1044-1092. DOI PMID
[43]	Umezawa T (2003). Diversity in lignan biosynthesis. Phytochem Rev 2, 371-390. DOI URL
[44]	Umezawa T, Davin LB, Lewis NG (1991). Formation of lignans (-)-secoisolariciresinol and (-)-matairesinol with Forsythia intermedia cell-free extracts. J Biol Chem 266, 10210-10217. PMID
[45]	van Fürden B, Humburg A, Fuss E (2005). Influence of methyl jasmonate on podophyllotoxinand 6-methoxypodophyllotoxin accumulation in Linum album cell suspension cultures. Plant Cell Rep 24, 312-317. DOI URL
[46]	von Heimendahl CBI, Schäfer KM, Eklund P, Sjöholm R, Schmidt TJ, Fuss E (2005). Pinoresinol-lariciresinol reductases with different stereospecificity from Linum album and Linum usitatissimum. Phytochemistry 66, 1254-1263. DOI PMID
[47]	Wada H, Kido T, Tanaka N, Murakami T, Saiki Y, Chen CM (1992). Chemical and chemotaxonomical studies of ferns. LXXXI. Characteristic lignans of Blechnaceous ferns. Chem Pharm Bull 40, 2099-2101. DOI URL
[48]	Wankhede DP, Biswas DK, Rajkumar S, Sinha AK (2013). Expressed sequence tags and molecular cloning and characterization of gene encoding pinoresinol/lariciresinol reductase from Podophyllum hexandrum. Protoplasma 250, 1239-1249. DOI URL
[49]	Wu YL, Xing DW, Ma GL, Dai XL, Gao LP, Xia T (2019). A variable loop involved in the substrate selectivity of pinoresinol/lariciresinol reductase from Camellia sinensis. Phytochemistry 162, 1-9. DOI URL
[50]	Xiao Y, Ji Q, Gao SH, Tan HX, Chen RB, Li Q, Chen JF, Yang YB, Zhang L, Wang ZT, Chen WS, Hu ZB (2015). Combined transcriptome and metabolite profiling reveals that IiPLR1 plays an important role in lariciresinol accumulation in Isatis indigotica. J Exp Bot 66, 6259-6271. DOI URL
[51]	Xiao Y, Shao K, Zhou JW, Wang L, Ma XQ, Wu D, Yang YB, Chen JF, Feng JX, Qiu S, Lv ZY, Zhang L, Zhang P, Chen WS (2021). Structure-based engineering of substrate specificity for pinoresinol-lariciresinol reductases. Nat Commun 12, 2828. DOI PMID
[52]	Zheng CJ, Zhang XW, Han T, Jiang YP, Tang JY, Brömme D, Qin LP (2014). Anti-inflammatory and anti-osteoporotic lignans from Vitex negundo seeds. Fitoterapia 93, 31-38. DOI URL
[53]	Zhou L, Yao GD, Lu LW, Song XY, Lin B, Wang XB, Huang XX, Song SJ (2018). Neolignans from red raspberry (Rubus idaeus L.) exhibit enantioselective neuroprotective effects against H₂O₂-induced oxidative injury in SH-SY5Y cells. J Agric Food Chem 66, 11390-11397. DOI URL

植物中松脂醇-落叶松脂素还原酶催化特征研究进展

Research Progress on Catalytic Characteristics of Pinoresinol-lariciresinol Reductase in Plants

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