Biochemical Properties and Phylogeny of Hydroxypyruvate Reductases from Methanotrophic Bacteria with Different C1-Assimilation Pathways

被引:10
作者
But, S. Y. [1 ]
Egorova, S. V. [2 ]
Khmelenina, V. N. [1 ]
Trotsenko, Y. A. [1 ,2 ]
机构
[1] Russian Acad Sci, Skryabin Inst Biochem & Physiol Microorganisms, Lab Methylotrophy, Pushchino 142290, Moscow Region, Russia
[2] Pushchino State Inst Nat Sci, Pushchino 142290, Moscow Region, Russia
基金
俄罗斯基础研究基金会;
关键词
hydroxypyruvate reductase; methanotrophs; serine cycle of carbon assimilation; METHYLOCOCCUS-CAPSULATUS BATH; D-GLYCERATE DEHYDROGENASE; PURIFICATION; NADPH; METHYLOTROPH;
D O I
10.1134/S0006297917110074
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
In the aerobic methanotrophic bacteria Methylomicrobium alcaliphilum 20Z, Methylococcus capsulatus Bath, and Methylosinus trichosporium OB3b, the biochemical properties of hydroxypyruvate reductase (Hpr), an indicator enzyme of the serine pathway for assimilation of reduced C-1-compounds, were comparatively analyzed. The recombinant Hpr obtained by cloning and heterologous expression of the hpr gene in Escherichia coli catalyzed NAD(P)H-dependent reduction of hydroxypyruvate or glyoxylate, but did not catalyze the reverse reactions of D-glycerate or glycolate oxidation. The absence of the glycerate dehydrogenase activity in the methanotrophic Hpr confirmed a key role of the enzyme in utilization of C-1-compounds via the serine cycle. The enzyme from Ms. trichosporium OB3b realizing the serine cycle as a sole assimilation pathway had much higher special activity and affinity in comparison to Hpr from Mm. alcaliphilum 20Z and Mc. capsulatus Bath assimilating carbon predominantly via the ribulose monophosphate (RuMP) cycle. The hpr gene was found as part of gene clusters coding the serine cycle enzymes in all sequenced methanotrophic genomes except the representatives of the Verrucomicrobia phylum. Phylogenetic analyses revealed two types of Hpr: (i) Hpr of methanotrophs belonging to the Gammaproteobacteria class, which use the serine cycle along with the RuMP cycle, as well as of non-methylotrophic bacteria belonging to the Alphaproteobacteria class; (ii) Hpr of methylotrophs from Alpha- and Betaproteobacteria classes that use only the serine cycle and of non-methylotrophic representatives of Betaproteobacteria. The putative role and origin of hydroxypyruvate reductase in methanotrophs are discussed.
引用
收藏
页码:1295 / 1303
页数:9
相关论文
共 20 条
[11]   Characterization of the pyrophosphate-dependent 6-phosphofructokinase from Methylococcus capsulatus Bath [J].
Reshetnikov, Alexander S. ;
Rozova, Olga N. ;
Khmelenina, Valentina N. ;
Mustakhimov, Ildar I. ;
Beschastny, Alexander P. ;
Murrell, J. Colin ;
Trotsenko, Yuri A. .
FEMS MICROBIOLOGY LETTERS, 2008, 288 (02) :202-210
[12]  
Rozova Olga N, 2015, Microorganisms, V3, P47, DOI 10.3390/microorganisms3010047
[13]  
Sambrook J., 2012, Molecular Cloning: A Laboratory Manual, V4th edn
[14]  
SHISHKINA V N, 1976, Mikrobiologiya, V45, P417
[15]   STABLE PATTERN FORMATION AND DETERMINATION OF MOLECULAR SIZE BY PORE-LIMIT ELECTROPHORESIS [J].
SLATER, GG .
ANALYTICAL CHEMISTRY, 1969, 41 (08) :1039-&
[16]   Metabolic aspects of aerobic obligate methanotrophy [J].
Trotsenko, Yuri A. ;
Murrell, John Colin .
ADVANCES IN APPLIED MICROBIOLOGY, VOL 63, 2008, 63 :183-229
[17]  
UTTING JM, 1975, J BIOL CHEM, V250, P5233
[18]   Genomic insights into methanotrophy:: The complete genome sequence of Methylococcus capsulatus (Bath) [J].
Ward, N ;
Larsen, O ;
Sakwa, J ;
Bruseth, L ;
Khouri, H ;
Durkin, AS ;
Dimitrov, G ;
Jiang, LX ;
Scanlan, D ;
Kang, KH ;
Lewis, M ;
Nelson, KE ;
Methé, B ;
Wu, M ;
Heidelberg, JF ;
Paulsen, IT ;
Fouts, D ;
Ravel, J ;
Tettelin, H ;
Ren, QH ;
Read, T ;
DeBoy, RT ;
Seshadri, R ;
Salzberg, SL ;
Jensen, HB ;
Birkeland, NK ;
Nelson, WC ;
Dodson, RJ ;
Grindhaug, SH ;
Holt, I ;
Eidhammer, I ;
Jonasen, I ;
Vanaken, S ;
Utterback, T ;
Feldblyum, TV ;
Fraser, CM ;
Lillehaug, JR ;
Eisen, JA .
PLOS BIOLOGY, 2004, 2 (10) :1616-1628
[19]  
Whittenbury R, 1980, P INT S MICR GROWTH, P181
[20]  
WILLIS JE, 1962, J BIOL CHEM, V237, P910