Easy and Efficient Permeabilization of Cyanobacteria for in vivo Enzyme Assays Using B-PER

Simon Matthe Erstad; Yumiko Sakuragi

Easy and Efficient Permeabilization of Cyanobacteria for in vivo Enzyme Assays Using B-PER

Simon Matthe Erstad, Yumiko Sakuragi

Section for Molecular Plant Biology

Abstract

Cyanobacteria are photosynthetic bacteria that thrive in diverse ecosystems and play major roles in the global carbon cycle. The abilities of cyanobacteria to fix atmospheric CO2 and to allocate the fixed carbons to chemicals and biofuels have attracted growing attentions as sustainable microbial cell factories. A better understanding of activities of enzymes involved in the central carbon metabolism might lead to increased product yields. Currently, cell-free lysates are widely used for the determination of intracellular enzyme activities. However, due to thick cell walls in cyanobacteria, lysis of cyanobacterial cells is inefficient and often laborious. The present protocol describes an easy and efficient method to permeabilize cyanobacterial cells, without lysing them, and direct usage of the permeabilized cells for the determination of metabolic enzyme activities in vivo.

Original language	English
Article number	Vol 8, Iss 01, Jan 05, 2018 DOI:10.21769/BioProtoc.2667
Journal	Bio-protocol
Pages (from-to)	1-10
Number of pages	10
ISSN	2331-8325
Publication status	Published - 5 Jan 2018

Cite this

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title = "Easy and Efficient Permeabilization of Cyanobacteria for in vivo Enzyme Assays Using B-PER",

abstract = "Cyanobacteria are photosynthetic bacteria that thrive in diverse ecosystems and play major roles in the global carbon cycle. The abilities of cyanobacteria to fix atmospheric CO2 and to allocate the fixed carbons to chemicals and biofuels have attracted growing attentions as sustainable microbial cell factories. A better understanding of activities of enzymes involved in the central carbon metabolism might lead to increased product yields. Currently, cell-free lysates are widely used for the determination of intracellular enzyme activities. However, due to thick cell walls in cyanobacteria, lysis of cyanobacterial cells is inefficient and often laborious. The present protocol describes an easy and efficient method to permeabilize cyanobacterial cells, without lysing them, and direct usage of the permeabilized cells for the determination of metabolic enzyme activities in vivo.",

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T1 - Easy and Efficient Permeabilization of Cyanobacteria for in vivo Enzyme Assays Using B-PER

AU - Erstad, Simon Matthe

AU - Sakuragi, Yumiko

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N2 - Cyanobacteria are photosynthetic bacteria that thrive in diverse ecosystems and play major roles in the global carbon cycle. The abilities of cyanobacteria to fix atmospheric CO2 and to allocate the fixed carbons to chemicals and biofuels have attracted growing attentions as sustainable microbial cell factories. A better understanding of activities of enzymes involved in the central carbon metabolism might lead to increased product yields. Currently, cell-free lysates are widely used for the determination of intracellular enzyme activities. However, due to thick cell walls in cyanobacteria, lysis of cyanobacterial cells is inefficient and often laborious. The present protocol describes an easy and efficient method to permeabilize cyanobacterial cells, without lysing them, and direct usage of the permeabilized cells for the determination of metabolic enzyme activities in vivo.

AB - Cyanobacteria are photosynthetic bacteria that thrive in diverse ecosystems and play major roles in the global carbon cycle. The abilities of cyanobacteria to fix atmospheric CO2 and to allocate the fixed carbons to chemicals and biofuels have attracted growing attentions as sustainable microbial cell factories. A better understanding of activities of enzymes involved in the central carbon metabolism might lead to increased product yields. Currently, cell-free lysates are widely used for the determination of intracellular enzyme activities. However, due to thick cell walls in cyanobacteria, lysis of cyanobacterial cells is inefficient and often laborious. The present protocol describes an easy and efficient method to permeabilize cyanobacterial cells, without lysing them, and direct usage of the permeabilized cells for the determination of metabolic enzyme activities in vivo.

M3 - Journal article

SN - 2331-8325

SP - 1

EP - 10

JO - Bio-protocol

JF - Bio-protocol

M1 - Vol 8, Iss 01, Jan 05, 2018 DOI:10.21769/BioProtoc.2667

ER -

Easy and Efficient Permeabilization of Cyanobacteria for in vivo Enzyme Assays Using B-PER

Abstract

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