What You Need to Know About BACTOSOMES® and Recombinant Enzymes’ Abilities for Research Purposes

When it comes to your research needs, BACTOSOMES® and recombinant enzymes are a vital part of the process. BioIVT is a leading distributor of BACTOSOMES® and partner with many researchers to reach their goals. But what do you need to know about BACTOSOMES®? In this blog we will discuss what BACTOSOMES® are and how you can apply them to your research needs.

What Are BACTOSOMES®?

Simply put, BACTOSOMES® are enzymes with bacterial membranes containing recombinant cytochrome P450 (CYP450) enzymes co-expressed with NADPH-cytochrome P450 (CRP) reductase. They are created by transforming E. coli bacteria into suitable DNA which causes them to express human or animal enzyme proteins. The reason this is done is because bacteria are ideal in this process because they don’t, themselves, express CYP450s. This results in low chances of background activity. 

Features of BACTOSOMES®

There are many features that make BACTOSOMES® ideal for research and testing. Some of those features include:

• Patented E. coli expression system

• Excellent batch-to-batch consistency

• Patented production method for native, or minimally modified CYP450’s

• Robust CYP450 activity levels

• Extensive batch-specific data (Kₘ, Vmax, linearity with time and enzyme, at Kₘ)

• Comprehensive selection, including modified-reductase choices and controls

• ISO9001:2009 compliant production and quality control

• 10-30 minute linearity over time

Due to excellent linearity over time, BACTOSOMES® allow longer incubations which generate better results. Since reductase and high reductase BACTOSOMES® contain high levels of CRP reductase, they exhibit limited linearity turnover with time which in turn creates higher activity. 

Benefits of BACTOSOMES®

The primary benefit here is that “regular” BACTOSOMES® is that you can use more CYP450 per milligram of the total protein. In metabolism identification studies, BACTOSOMES® would be preferred due to their higher concentrations of CYP450. Research has demonstrated that CYP450s expressed in E. coli shows more enzymatic activity than that compared to the same enzymes in insect cells. In addition, BACTOSOMES® demonstrate greater turnover numbers when compared to insect cells. These results show that human CYP450 expressed in E. coli are commonly more active than the CYP450 found in insect cells.

Other benefits of BACTOSOMES® include:

•  BACTOSOMES® utilize native P450 and better represent human enzymes

• Available with and without b5

• Different levels of reductase allow for high and low activities

• Reductase BACTOSOMES® are optimized for metabolism

• Low Reductase  BACTOSOMES® developed specifically for inhibition assays

•  BACTOSOMES® exhibit the best linearity over time compared to competitor CYP450s

• Least expensive commercial option

• Extensive panel of CYP450s and Phase II expressed enzymes available

• Only commercial source for recombinant dog CYP450s

Recommendations for Product Application

BACTOSOMES® are extraordinarily effective in test systems requiring high levels of individualized enzymatic activities. Reaction phenotyping studies are the most common applications for recombinant enzymes. Additionally, these enzymes are useful in screening programs for enzyme inhibition and in metabolite synthesis.

Research Applications and How BACTOSOMES® Fit in In Vitro ADME Research Programs

Recombinant CYPs are regularly used for ADME-Tox studies as in vitro tools. This occurs mostly in enzyme inhibition screening, reaction phenotyping, and clearance/metabolite ID.

Custom Synthesis Capabilities

At BioIVT, our system has demonstrated effectiveness for a broad range of enzymes and we have routinely assembled custom synthesis projects. With our process, projects are scalable from 100 milliliter shake flasks to 80 liter fermenters.

The first step in any custom synthesis project is the assessment of the protein of interest to determine the best transfection approach feasible.

Our Enzyme Catalog 

Human Recombinant Enzymes

BioIVT boasts an impressive catalog of human and animal BACTOSOMES® as well as purified enzymes, including CYPs commonly investigated in drug discovery programs. These include:

Human BACTOSOMES® Enzymes

Human BACTOSOMES® Enzymes are specimens available in 1 nmol vials and are shipped and stored at -80℃.

Human EasyCYP™ BACTOSOMES® Enzymes

Human EasyCYP™ BACTOSOMES® Enzymes are specimens available in 0.5 nmol vials at a protein concentration of 10 mg/mL and are shipped and stored -80℃.

Human Purified CYP Enzymes

Human Purified CYP Enzymes are customized to customer specifications.

Animal Recombinant Enzymes

Canine BACTOSOMES® Enzymes

Canine BACTOSOMES® Enzymes are specimens available in 1 nmol vials and are shipped and stored at -80℃.

Monkey BACTOSOMES® Enzymes

Monkey BACTOSOMES® Enzymes are specimens available in 1 nmol vials and are shipped and stored at -80℃.

Mouse BACTOSOMES® Enzymes

Mouse BACTOSOMES® Enzymes are specimens available in 1 nmol vials and are shipped and stored at -80℃.

Pig BACTOSOMES® Enzymes

Pig BACTOSOMES® Enzymes are specimens available in 1 nmol vials and are shipped and stored at -80℃.

Rat BACTOSOMES® Enzymes

Rat BACTOSOMES® Enzymes are specimens available in 1 nmol vials and are shipped and stored at -80℃.

Non-CYP Recombinant Enzymes

BioIVT boasts an impressive catalog of non-CYP recombinant metabolizing enzymes. These enzymes are primarily used for drug discovery and preclinical development for applications in studies requiring extensive individual enzymatic activities. In vitro models using recombinant enzymes are the simplest test system available in studying interactions and are effective in identifying enzymes involved in the metabolism of a drug candidate. These enzymes include:

Human Aldehyde Dehydrogenase (ALDH)

Human Aldehyde Dehydrogenase (ALDH) or 1A1 is expressed in E. coli (10 mg/mL) in 10 mM sodium phosphate (pH 7.4) containing 140 mM NaCl. Each vial of human ALDH recombinant enzyme contains 1 mg of cytosolic protein.

Human Aldehyde Oxidase (AOX)

Human Aldehyde Oxidase (AOX) is expressed in E. coli (10 mg/mL) in 25 mM potassium phosphate (pH 7.4) containing 0.1 mM EDTA. Each vial of human AOX recombinant enzyme contains 2 mg of cytosolic protein.

Human Carboxylesterase (CES)

Human Carboxylesterase (CES) comes in two types. Human Carboxylesterase 1 (CES1) and Human Carboxylesterase 2 (CES2) are expressed in E. coli (10 mg/mL) in 50 mM Tris-acetate (pH 7.6), 250 mM sucrose, 0.25 mM EDTA. Each vial of human CES recombinant enzyme contains 1 mg of membrane protein.

Human Glutathione S-transferase (GST)

Human Glutathione S-transferase (GST) comes in three types. Human Glutathione S-Transferase A1 (GSTA1), Human Glutathione S-Transferase M1 (GSTM1), and Human Glutathione S-Transferase P1 (GSTP1) recombinant enzymes are expressed in E. coli (10 mg/mL) and stored in 10 mM sodium phosphate (pH 7.4) with 140 mM NaCl. Each vial of human GST recombinant enzyme contains 1 mg of cytosolic protein.

Human Monoamine Oxidase (MAO)

Human Monoamine Oxidase (MAO) comes in two types. Human Monoamine Oxidase A (MAOA) and Human Monoamine Oxidase B (MAOB) are expressed in insect cells (5uµg/mL) in 10 mM sodium phosphate (pH 7.4), 140 mM NaCl, 20% (v/v) glycerol. Each vial of MAO recombinant enzyme contains 2.5 mg protein.

Human Sulfotransferase (SULT)

Human Sulfotransferase (SULT) comes in a variety of types. Human sulfotransferases (SULTs) 1A1*1, SULT1B1, SULT1E1, SULT2A1, SULT1A1*2, SULT1A2, SULT1C2, and SULT1C4 recombinant enzymes are expressed in E. coli (10 mg/mL) in 10 mM sodium phosphate (pH 7.4), 140 mM NaCl. Each vial of human SULT recombinant enzyme contains 1 mg cytosolic protein.

Human UDP-glucuronosyltransferase (UGT)

Human UDP-glucuronosyltransferase (UGT) comes in a variety of types. Human UDP-glucuronosyltransferases (UGTs) 1A1, UGT1A3, UGT1A4, UGT1A6, UGT1A7, UGT1A8, UGT1A9, UGT1A10, UGT2B7, UGT2B17 are expressed in Sf9 insect cells (5 mg/mL) in 25mM Tris-acetate (pH 7.6), 10% (v/v) glycerol, 1mM EDTA. Each vial of UGT recombinant enzyme contains 2.5 mg protein.