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. 2019 Nov 20;24(23):4203.
doi: 10.3390/molecules24234203.

Effects of GGT and C-S Lyase on the Generation of Endogenous Formaldehyde in Lentinula edodes at Different Growth Stages

Xiaoyu Lei  1 , Shuangshuang Gao  1 , Xi Feng  2 , Zhicheng Huang  1 , Yinbing Bian  3 , Wen Huang  1 , Ying Liu  1
Affiliations

Effects of GGT and C-S Lyase on the Generation of Endogenous Formaldehyde in Lentinula edodes at Different Growth Stages

Xiaoyu Lei et al. Molecules. .

Abstract

Endogenous formaldehyde is generated as a normal metabolite via bio-catalysis of γ-glutamyl transpeptidase (GGT) and L-cysteine sulfoxide lyase (C-S lyase) during the growth and development of Lentinula edodes. In this study, we investigated the mRNA and protein expression levels, the activities of GGT and C-S lyase, and the endogenous formaldehyde content in L. edodes at different growth stages. With the growth of L. edodes, a decrease was found in the mRNA and protein expression levels of GGT, while an increase was observed in the mRNA and protein expression levels of C-S lyase as well as the activities of GGT and C-S lyase. Our results revealed for the first time a positive relationship of formaldehyde content with the expression levels of Csl (encoding Lecsl) and Lecsl (C-S lyase protein of Lentinula edodes) as well as the enzyme activities of C-S lyase and GGT during the growth of L. edodes. This research provided a molecular basis for understanding and controlling the endogenous formaldehyde formation in Lentinula edodes in the process of growth.

Keywords: C-S lyase; GGT; Lentinula edodes; endogenous formaldehyde; expression levels.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Proposed pathway for the generation of sulfurous flavor compounds and endogenous formaldehyde in Lentinula edodes.
Figure 2
Figure 2
Expression of Ggtl and Csl during fruit-body development of Lentinula edodes strain W1. Relative expression during five growth stages, M, mycelia; G, grey; YFB, young fruiting body; IFB, immature fruiting body; MFB, mature fruiting body. Transcript levels of Ggtl (black bars) and Csl (gray bars) were determined by real-time quantitative RT-PCR analysis and normalized against Actinl. The expressions during the M stage were taken as 1. Error bars indicate standard deviation for three independent experiments. *p < 0.05, ANOVA tests by Duncan’s indicate significant differences.
Figure 3
Figure 3
(A) Protein levels of Leggt at five stages of growth. (B) Protein levels of Lecsl at five stages of growth. (C) Relative expression of Leggt and Lecsl during five growth stages. β-actin protein was used as loading control, and the expressions during the M stage were taken as 100%. M, mycelia; G, grey; YFB, young fruiting body; IFB, immature fruiting body; MFB, mature fruiting body. Error bars indicate standard deviation for three independent experiments. *p < 0.05, ANOVA tests by Duncan’s indicate significant differences.
Figure 4
Figure 4
(A) The specific activity of GGT at the five growth stages. The reaction mixture containing the enzyme and the GPNA substrate was analyzed under standard conditions, and the residual activity was calculated. (B) The specific activity of C-S lyase at the five growth stages. The reaction mixture containing the enzyme and S-ethyl-L-cysteine sulfoxide substrate was analyzed under standard conditions, and the residual activity was calculated. M, mycelia; G, grey; YFB, young fruiting body; IFB, immature fruiting body; MFB, mature fruiting body. Error bars indicate standard deviation for three independent experiments. *p < 0.05, ANOVA tests by Duncan’s indicate significant differences.
Figure 5
Figure 5
Endogenous formaldehyde content of L. edodes strain W1 at different growth stages. M, mycelia; G, grey; YFB, young fruiting body; IFB, immature fruiting body; MFB, mature fruiting body. Error bars indicate standard deviation for three independent experiments. *p < 0.05, ANOVA tests by Duncan’s indicate significant differences.
Figure 6
Figure 6
Five growth stages of L. edodes strain W1. (A) Mycelia. (B) Grey (5–10 mm in cap diameter). (C) Young fruiting body (15–20 mm in cap diameter). (D) Immature fruiting body (with partial veil not ruptured). (E) Mature fruiting body (with partial veil entirely ruptured).
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