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DUF538 protein superfamily is predicted to be chlorophyll hydrolyzing enzymes in plants

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Abstract

The possible hydrolytic activity towards chlorophyll molecules was predicted for DUF538 protein superfamily in plants. It was examined by using computational as well as experimental tools including in vitro chlorophyll degradation, antioxidant compounds production and in vivo real-time gene expression tests. Comparison of the computational data with the experimental results indicated that DUF538 proteins might be chlorophyll hydrolyzing enzyme (most probably carboxyesterase) which degrade chlorophyll molecules (66 % per 12 hrs) to produce new compounds (1.8 fold per 12 hrs) with antioxidant properties. The relevance of DUF538 gene expression level with the chlorophyll contents (2.8 fold increase per chlorophyll content of 50 %) of the drought-stressed leaves showed that chlorophyll degradation by DUF538 is most probably induced in response to stress stimuli. Despite membranous chlorophyll catabolic pathways, DUF538-dependent reactions is predicted to be occurred in the cytosol of the under stressed plants. We addressed as to whether chlorophyll breakdown to antioxidant compounds by DUF538 is a defense mechanism of plants against stress stimuli, in vivo? This question is going to be investigated in our next research project.

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Abbreviations

DUF538:

Domain of unknown function

NMR:

nuclear magnetic resonance

BPI:

Bactericidal permeability increasing

WSCP1:

Water soluble chlorophyll binding protein 1

LPS:

Lipopolysaccharide

ROS:

Reactive oxygen species

MBP:

Maltose-binding protein

IPTG:

Isopropyl β-D-thiogalactopyranoside

EDTA:

Ethylenediaminetetracetic

BME:

ß-mercaptoethanol

BSA:

Bovine serum albumin

NBT:

Nitro blue tetrazolium

BCIP:

Bromo chloro indolyl phosphate

DMF:

Dimethyl formamide

FRAP:

Ferric reducing antioxidant power

TPTZ:

Tripyridyl triazine

RT-PCR:

Reverse transcriptase-polymerase chain reaction

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Acknowledgments

The author of this paper is thankful to the Research Institute for Fundamental Sciences (RIFS), University of Tabriz for the financial support.

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Authors and Affiliations

  1. Research Institute for Fundamental Sciences (RIFS), University of Tabriz, Tabriz, Iran

    Ashraf Gholizadeh

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  1. Ashraf Gholizadeh
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Correspondence to Ashraf Gholizadeh.

Appendix

Appendix

Figure 6

Fig. 6
figure 6

Analysis of DUF538 distribution in nature. Ortholog cluster was analyzed by OC server at www.genome.ad.jp. Organisms sequence include Eudicots, Monocots, Basal Magnoliophyta, Ferns and Mosses

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Gholizadeh, A. DUF538 protein superfamily is predicted to be chlorophyll hydrolyzing enzymes in plants. Physiol Mol Biol Plants 22, 77–85 (2016). https://doi.org/10.1007/s12298-015-0331-1

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