Multi-step enzyme reaction in an Immobilized Enzyme Microreactor (IEMR)
The application of biocatalyst in microfluidic systems has attracted huge attention in recent years due to their potential for creating microscale analytical tools with small volumes, high-throughput and attractive prospect for mass production. This system can be integrated with the multi-step enz...
| Main Authors: | , |
|---|---|
| Format: | Conference or Workshop Item |
| Language: | English |
| Published: |
Kulliyah of Engineering, International Islamic University Malaysia
2016
|
| Subjects: | |
| Online Access: | http://irep.iium.edu.my/52183/ http://irep.iium.edu.my/52183/ http://irep.iium.edu.my/52183/1/52183.pdf |
| Summary: | The application of biocatalyst in microfluidic systems has attracted huge attention in recent years due to their
potential for creating microscale analytical tools with small volumes, high-throughput and attractive prospect
for mass production. This system can be integrated with the multi-step enzyme reaction to synthesize complex
organic compounds such as a single diastereoisomer of 2-amino-1,3,4-butanetriol (ABT). ABT is an
interesting compound, commonly used as the building block in several pharmaceutical syntheses. In this
work we study a de novo designed pathway for the production of ABT using IEMRs, which is based on the
reversible attachment of His6-tagged enzymes via Ni-NTA agarose beads that are packed into tubes to enable
continuous flow of multi-step enzyme reactions. The synthesis of ABT was demonstrated in a three-step
enzyme reaction using two enzymes; transketolase and transaminase. In the first part of the three-step
reaction, His6-TAm catalysed the conversion of the substrates, serine (SER) and glycoaldehyde (GA), to form
lithium-hydroxypyruvate (HPA). The product of the first reaction was then continuously fed to the second
IEMR containing the immobilized His6-TK, which catalysed the reaction between HPA and GA into
erythrulose (ERY). The multi-step reaction was completed with the amination of ERY by the second His6-
TAm to produce the final target product, ABT. |
|---|