MDH E. coli

Malate Dehydrogenase Recombinant
Cat. No.
BT11520
Source
E.coli.
Synonyms
Malate dehydrogenase cytoplasmic, EC 1.1.1.37, Cytosolic malate dehydrogenase, MDHA, MOR2, MDH-s, MGC:1375, MDH1.
Appearance
Sterile Filtered colorless solution.
Purity
Greater than 95% as determined by SDS-PAGE.
Usage
THE BioTek's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
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Description

MDH Recombinant produced in E. coli is a single polypeptide chain containing 336 amino acids (1-312) and having a molecular mass of 34.9kDa.
MDH is fused to a 24 amino acid His-tag at N-terminus & purified by proprietary chromatographic techniques.

Product Specs

Introduction
Malate dehydrogenase (also known as MDH or EC 1.1.1.37) is an enzyme that plays a crucial role in the citric acid cycle and gluconeogenesis. It catalyzes the reversible conversion of malate to oxaloacetate, a key intermediate in these metabolic pathways. This enzyme utilizes NAD+ as a cofactor. It's important to differentiate MDH from malic enzyme, as they catalyze distinct reactions.
Description
This recombinant MDH, produced in E. coli, is a single polypeptide chain consisting of 336 amino acids (with the first 312 amino acids forming the MDH sequence). It has a molecular weight of 34.9 kDa. For purification purposes, a 24 amino acid His-tag is attached to the N-terminus. The protein is purified using proprietary chromatographic techniques.
Physical Appearance
Clear, colorless solution that has been sterilized by filtration.
Formulation
The MDH is supplied in a solution at a concentration of 1mg/ml. The solution is buffered with 20mM Tris-HCl at pH 8.0 and contains 50mM Nacl, 1mM DTT, and 10% glycerol for stability.
Stability
For short-term storage (2-4 weeks), the MDH solution can be stored at 4°C. For long-term storage, it is recommended to store the solution frozen at -20°C. Adding a carrier protein like HSA or BSA (0.1%) can enhance stability during long-term storage. To maintain the enzyme's activity, avoid repeated freezing and thawing cycles.
Purity
The purity of MDH is greater than 95%, as determined by SDS-PAGE analysis.
Synonyms
Malate dehydrogenase cytoplasmic, EC 1.1.1.37, Cytosolic malate dehydrogenase, MDHA, MOR2, MDH-s, MGC:1375, MDH1.
Source
E.coli.
Amino Acid Sequence
MGSSHHHHHH SSGLVPRGSH MGSHMKVAVL GAAGGIGQAL ALLLKTQLPS GSELSLYDIA PVTPGVAVDL SHIPTAVKIK GFSGEDATPA LEGADVVLIS AGVARKPGMD RSDLFNVNAG IVKNLVQQVA KTCPKACIGI ITNPVNTTVA IAAEVLKKAG VYDKNKLFGV TTLDIIRSNT FVAELKGKQP GEVEVPVIGG HSGVTILPLL SQVPGVSFTE QEVADLTKRI QNAGTEVVEA KAGGGSATLS MGQAAARFGL SLVRALQGEQ GVVECAYVEG DGQYARFFSQ PLLLGKNGVE ERKSIGTLSA FEQNALEGML DTLKKDIALG EEFVNK

Product Science Overview

Introduction

Malate dehydrogenase (MDH) is a crucial enzyme in cellular metabolism, playing a vital role in the citric acid cycle (Krebs cycle). It catalyzes the reversible conversion of malate to oxaloacetate while reducing NAD+ to NADH. This enzyme is ubiquitous, found in all kingdoms of life, and is essential for energy production and various metabolic pathways .

Structure and Isoforms

MDH exists in multiple isoforms, primarily cytosolic MDH (cyMDH) and mitochondrial MDH (mMDH). These isoforms are localized in different cellular compartments and have distinct physiological roles. The cytosolic form is involved in the malate-aspartate shuttle, which transfers reducing equivalents across the mitochondrial membrane, while the mitochondrial form is directly involved in the citric acid cycle .

Recombinant MDH

Recombinant MDH refers to the enzyme produced through recombinant DNA technology. This involves inserting the gene encoding MDH into a suitable expression system, such as bacteria, yeast, or mammalian cells, to produce the enzyme in large quantities. Recombinant MDH is used extensively in research and industrial applications due to its high purity and activity.

Functional Characterization

The functional characterization of recombinant MDH involves studying its catalytic properties, substrate specificity, and kinetic parameters. MDH catalyzes the dehydrogenation of malic acid to generate oxaloacetic acid, accompanied by the reduction of NAD+ to NADH. This reaction is crucial for maintaining the redox balance within the cell and is involved in various metabolic pathways, including gluconeogenesis, amino acid metabolism, and lipid biosynthesis .

Applications

Recombinant MDH has several applications in biotechnology and research:

  1. Metabolic Engineering: It is used to engineer metabolic pathways in microorganisms for the production of biofuels and biochemicals.
  2. Diagnostic Kits: MDH is a component of diagnostic assays for measuring levels of NADH and NAD+ in clinical samples.
  3. Structural Studies: Recombinant MDH is used in crystallography and molecular dynamics simulations to study enzyme structure and function .

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