硫对酵母生物富铬过程中铬胁迫的缓解作用

doi:10.3864/j.issn.0578-1752.2019.06.011

Abstract

Abstract:

【Objective】The objective of this study was to investigate the antioxidative mechanism and role of sulphur during chromium (Ⅲ) enrichment by Saccharomyces cerevisiae. The mechanisms of alleviation chromium (Ⅲ) toxicity against yeast by sulphur were revealed. 【Method】Saccharomyces cerevisiae YSI-3.7 was used in this study. Various incubation conditions were investigated, such as various concentrations Cr(Ⅲ) and sulfate. And the corresponding biomass, total chromium content, organic chromium content and oxidative stress markers (including malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and so on), were measured and analyzed. 【Result】 Low Cr(Ⅲ) concentration (0-200 μg?mL ^-1) could stimulate the growth of yeast, while high Cr(Ⅲ) concentration (>500 μg?mL ^-1) inhibited its growth. After Saccharomyces cerevisiae YSI-3.7 was incubated with 500 μg?mL ^-1 Cr(Ⅲ) for 44 h, the content of organic Cr in yeast was found to be 725.55±55.08 μg?g ^-1 DCW and that of total Cr was 1255.53±43.75 μg?g ^-1 DCW. After Saccharomyces cerevisiae YSI-3.7 was incubated with 800 μg?mL ^-1 Cr(Ⅲ) for 44 h, the content of organic Cr in yeast was found to be 536.25±36.89 μg?g ^-1 DCW and that of total Cr was 1812.22±38.24 μg?g ^-1 DCW. The content of MDA increased (from 11.83 nmol?mL ^-1 to 18.04 nmol?mL ^-1) with the increasement of Cr(Ⅲ) concentration (0-800 μg?mL ^-1), while the activity of SOD and CAT decreased. The content of GSH, total sulfhydryl and T-AOC increased at lower Cr(Ⅲ) concentration (≤500 μg?mL ^-1), and decreased at the high concentration (800 μg?mL ^-1). The supplementation of 1 mmol?L ^-1 Na₂SO₃during incubation could alleviate the stress of Cr(Ⅲ) against yeast. The protein content increased and MDA content decreased (12.83%) with the addition of 1 mmol?L ^-1 Na₂SO₃ during incubation. The activity of CAT was almost unaffected. The activity of SOD was increased to 4.41%. GSH, T-AOC and GSH-Px content increased to 28.83%, 14.29% and 18.80%, respectively. 【Conclusion】During the Cr(Ⅲ) bio-enrichment process by yeast, Cr(Ⅲ) stress could aggravate the lipid peroxidation of cell membrane. At low Cr(Ⅲ) concentration(0-500 μg?mL ^-1), yeast could protect itself from this stress by its own antioxidant enzymes, among which glutathione and its related enzymes played an important role. At high concentration of Cr(Ⅲ) (800 μg?mL ^-1) , the degree of membrane lipid peroxidation was aggravated and the yeast’s own antioxidant capacity was not enough to protect itself from Cr(Ⅲ) stress. Supplementation of S (1 mmol?L ^-1 Na₂SO₃) could mitigate membrane lipid peroxidation caused by Cr(Ⅲ)by improving SOD activity, GSH, T-AOC and GSH-Px content in yeast, improving the antioxidant capacity of yeast itself and Cr(Ⅲ) bio-enrichment by yeast.

Key words: Cr(Ⅲ);, S, Saccharomyces cerevisiae, GTF, oxidative stress

LI HanTong,JIA ChengLi,ZHANG ShuWen,LU Jing,PANG XiaoYang,LIU Lu,LÜ JiaPing. Chromium (III) Stress Alleviation by Sulfur Compounds During Chromium Bio-enrichment by Saccharomyces cerevisiae[J].Scientia Agricultura Sinica, 2019, 52(6): 1078-1089.

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	Cr（Ⅲ） (μg?mL^-1)
	0	200	500	800
生物量 Biomass (g/100 mL)	1.40±0.07	1.06±0.01	0.97±0.05	0.45±0.02
有机铬 Organic chromium (μg·g^-1DCW)	0	224.74±6.41	725.55±55.08	536.25±36.89
总铬 Total chromium (μg·g^-1DCW)	0	409.04±12.65	1255.53±43.75	1812.22±38.24
有机铬/总铬 Organic/total chromium (%)	0	54.94±1.90	57.79±2.45	29.59±2.48

检测指标 Index	Cr（Ⅲ）浓度 Cr（Ⅲ） concentration (μg?mL^-1)
检测指标 Index	0	200	500	800
丙二醛 MDA (nmol?mL^-1)	11.83±0.38	15.54±0.41	16.91±0.33	18.04±0.44
超氧化物岐化酶 SOD (U?mg^-1prot)	8.61±0.19	8.28±0.11	7.93±0.07	7.35±0.21
过氧化氢酶 CAT (U?mg^-1prot)	9.09±0.28	7.83±0.14	4.72±0.09	3.64±0.03
还原型谷胱甘肽 GSH (μmol?g^-1 prot)	31.42±1.54	40.22±1.38	48.52±2.01	43.84±1.94
氧化型谷胱甘肽 GSSG (μmol?g^-1 prot)	7.86±0.21	9.56±0.32	12.38±0.52	15.46±0.61
谷胱甘肽氧化酶 GSH-Px (U?g^-1 prot)	1044.51±10.22	1274.32±5.89	763.01±3.21	534.68±6.76
总巯基 -SH (μg?g^-1 prot)	38.51±1.14	53.44±2.71	59.05±2.98	35.74±1.97
总抗氧化能力 T-AOC (U?g^-1 prot)	0.8±0.21	0.85±0.19	1.61±0.32	0.35±0.01

含硫化合物 S compound	浓度 Concentration (mmol?L^-1)	生物量 Biomass (g/100 mL)	有机铬 Organic chromium (μg·g^-1·DCW)	总铬 Total chromium (μg·g^-1·DCW)	有机铬率 Percentage of organic chromium (%)
0	对照组 Control	1.002±0.1	755.63±4.23	1431.93±10.12	52.77±1.24
Na₂SO₃	0.5	0.92±0.07	1198.16±9.43	1578.21±11.27	75.92±3.75
	1	0.85±0.08	1607.02±6.78	1876.32±9.43	85.65±2.36
	5	0.60±0.04	2052.43±10.58	2755.04±11.38	74.50±3.57
	10	0.49±0.03	609.58±4.98	1746.95±8.79	34.84±1.98
	15	0.32±0.02	477.39±2.75	740.18±8.02	64.50±1.63
Na₂S	0.5	1.005±0.25	895.98±5.72	1601.45±5.98	55.95±1.48
	1	1.08±0.07	1261.16±6.27	1868.74±9.98	67.75±2.32
	5	0.781±0.03	691.63±3.61	2302.31±11.54	30.04±1.03
	10	0.69±0.08	613.83±2.88	3516.62±21.32	17.46±0.98
	15	0.15±0.04	318.70±1.02	3834.02±20.98	8.31±0.45
（NH₄）₂SO₃	0.5	0.996±0.05	721.01±2.79	2101.45±14.32	34.31±1.74
	1	0.942±0.04	861.12±5.97	2868.14±16.45	30.02±1.05
	5	0.276±0.02	542.19±4.34	3000.21±18.91	18.07±1.13
	10	0.121±0.02	301.21±2.83	1812.62±13.69	16.62±0.45
	15	0.085±0.01	100.32±1.56	1367.02±9.13	7.33±0.41

Chromium (III) Stress Alleviation by Sulfur Compounds During Chromium Bio-enrichment by Saccharomyces cerevisiae

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