1.Deficiency of succinic dehydrogenase or succinyl-coA synthetase enhances the production of 5-aminolevulinic acid in recombinant Escherichia coli.
Wei PU ; Jiuzhou CHEN ; Cunmin SUN ; Ning CHEN ; Jibin SUN ; Ping ZHENG ; Yanhe MA
Chinese Journal of Biotechnology 2013;29(10):1494-1503
5-aminolevulinic acid (ALA), a precursor for biosynthesis of pyrrole compounds in living organisms, has been widely used in agriculture and medical photodynamics therapy and is regarded as a promising value-added bio-based chemical. In the previous investigations on ALA production with recombinant Escherichia coli expressing heterogenous C4 pathway gene, LB media supplemented with glucose and ALA precursors succinate and glycine is widely used, leading to high production cost. Succinate participates in ALA biosynthesis in a form of succinyl-CoA. In this study, genes involved in succinyl-CoA consumption, sdhAB (encoding succinic dehydrogenase) or sucCD (encoding succinyl-CoA synthetase) of E. coli MG1655 was knocked out and tested for ALA accumulation. In comparison with the recombinant E. coli strain expressing heterogenous ALA synthetase, the sdhAB- or sucCD-deficient strain accumulate 25.59% and 12.40%, respectively, more ALA in a 5 L fermentor using a defined synthetic medium with glucose as main carbon source and without supplementation of succinate, providing a novel cost-effective approach for industrial production of ALA.
Aminolevulinic Acid
;
metabolism
;
Escherichia coli
;
enzymology
;
genetics
;
metabolism
;
Industrial Microbiology
;
methods
;
Metabolic Engineering
;
methods
;
Recombinant Proteins
;
genetics
;
metabolism
;
Succinate Dehydrogenase
;
genetics
;
metabolism
;
Succinate-CoA Ligases
;
genetics
;
metabolism
2.Analysis of gene variant in an infant with succinic semialdehyde dehydrogenase deficiency.
Dandan YAN ; Xiaowei XU ; Xuetao WANG ; Xinjie ZHANG ; Xiufang ZHI ; Hong WANG ; Yuqing ZHANG ; Jianbo SHU
Chinese Journal of Medical Genetics 2022;39(2):216-221
OBJECTIVE:
To explore the genetic basis for a child with succinate semialdehyde dehydrogenase deficiency.
METHODS:
Peripheral blood samples of the proband and his parents were collected and subjected to Sanger sequencing. High-throughput sequencing was used to verify the gene variants. Bioinformatic software was used to analyze the pathogenicity of the variant sites.
RESULTS:
Sanger sequencing showed that the proband carried a homozygous c.1529C>T (p.S510F) variant of the ALDH5A1 gene, for which his mother was a carrier. The same variant was not detected in his father. However, high-throughput sequencing revealed that the child and his father both had a deletion of ALDH5A1 gene fragment (chr6: 24 403 265-24 566 986).
CONCLUSION
The c.1529C>T variant of the ALDH5A1 gene and deletion of ALDH5A1 gene fragment probably underlay the disease in the child. High-throughput sequencing can detect site variation as well as deletion of gene fragment, which has enabled genetic diagnosis and counseling for the family.
Amino Acid Metabolism, Inborn Errors/genetics*
;
Child
;
Developmental Disabilities
;
Humans
;
Infant
;
Mutation
;
Succinate-Semialdehyde Dehydrogenase/genetics*
3.Change in gastrocnemius dystrophin and metabolic enzymes and increase in high-speed exhaustive time induced by hypoxic training in rats.
Yu-Ming XU ; Jun-Ping LI ; Rui-Yuan WANG
Acta Physiologica Sinica 2012;64(4):455-462
The aim of the present study was to explore the changes and roles of dystrophin and membrane permeability in hypoxic training. Seventy-two 8-week-old Sprague Dawley (SD) rats were randomly divided into 4 groups, normoxic non-train (NC), normoxic train (NT), hypoxic non-train (HC), and hypoxic train (HT) groups. The rats of each group were randomly divided into three subgroups, non-exhaustive, low-speed exhaustive test and high-speed exhaustive test subgroups. Rats in hypoxia groups lived and were trained in a condition of 12.7% oxygen concentration (equal to the 4 300 m altitude). NT and HT groups received 4 weeks of training exercise. Then the rats in all non-exhaustive subgroups were sacrificed, and gastrocnemii were sampled for the measurements of lactate dehydrogenase (LDH), succinatedehydrogenase (SDH), malate dehydrogenase (MDH) activities. Moreover, serum LDH activity was analyzed. Low-speed exhaustive test and high-speed exhaustive test subgroups received exhaustive tests with 20 (71% VO2max) and 30 m/min speed (86% VO2max), respectively, and their exhaustive times were recorded. The results showed that, compared with normoxic groups, the weights in hypoxia groups exhibited slower increase. The level of dystrophin in HT group without exhaustion test didn't change significantly. The muscle MDH activities were markedly affected by the different oxygen concentration, training and their interaction (P < 0.05), whereas the muscle LDH activities were only affected by the different oxygen concentration (P < 0.05). Serum LDH activities were affected by the interaction of the different oxygen concentration and training (P < 0.05), showing decreased muscle LDH and increased blood LDH activities. The exhaustion time were markedly affected by the different test speed, training and their interaction (P < 0.05), and also affected by the interaction of the different oxygen concentration and training (P < 0.05), but didn't affected by oxygen concentration. The exhaustive time of HT high-speed exhaustive test subgroup was more than NT high-speed exhaustive test subgroup in 30 m/min exhaustion test. Compared with NT high-speed exhaustive test subgroup, HT high-speed exhaustive test subgroup had an earlier fatigue in the test, but had a rapid recovery. These results suggested that hypoxic training can effectively increase the rats' high-speed exhaustive time. The mechanism may be related to an increase in serum LDH caused by the increased membrane permeability after hypoxic training.
Altitude
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Animals
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Dystrophin
;
metabolism
;
Fatigue
;
Hypoxia
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L-Lactate Dehydrogenase
;
metabolism
;
Malate Dehydrogenase
;
metabolism
;
Muscle, Skeletal
;
enzymology
;
Physical Conditioning, Animal
;
Rats
;
Rats, Sprague-Dawley
;
Succinate Dehydrogenase
;
metabolism
4.Low expression of succinate dehydrogenase subunit B in locally recurrent nasopharyngeal carcinoma and implication for prognosis.
Sen LIN ; Zhijian DAI ; Chongxi CHEN ; Shenghua PAN ; Zhisu LIAO
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2015;50(1):38-42
OBJECTIVETo study the expression of succinate dehydrogenase subunit B (SDHB) in the tissues of locally recurrent nasopharyngeal carcinoma (rNPC) and the correlation with the clinicopathological factors and prognosis of rNPC.
METHODSImmunohistochemistry was used to detect the expression of SDHB in the tissues of primary and locally recurrent nasopharyngeal carcinoma. The relationship between SDHB expression and clinicopathological features was analyzed using the Chi-square test, and Kaplan-Meier method and Log-rank test were used for survival analysis. The independent prognostic factors of rNPC were analyzed by Cox regression model.
RESULTSLow SDHB expression was showed in 76.5% (39/51) of the patients with rNPC, significantly higher than 57.1% (24/42) of primary nasopharyngeal carcinoma (χ(2) = 4.098, P < 0.05). Low expression of SDHB strongly was correlated with T classification, clinical stage and cranial nerve palsy. Patients with low SDHB expression had a shorter survival time and a lower 3 or 5 year survival rate compared to the patients with high SDHB expression. Multivariate analysis showed that low SDHB expression was an independent predictor for overall survival of patients with rNPC (P < 0.05).
CONCLUSIONThe low SDHB expression is an independent indicator for poor prognosis of rNPC and may play an important role in the recurrence of rNPC.
Biomarkers, Tumor ; metabolism ; Carcinoma ; Humans ; Immunohistochemistry ; Multivariate Analysis ; Nasopharyngeal Neoplasms ; diagnosis ; metabolism ; Prognosis ; Succinate Dehydrogenase ; metabolism ; Survival Analysis
6.Effects of cocaine on activities of ATPase, LDH and SDH in mouse splenocytes.
Wen-ping SUN ; Yan-xu LU ; Xiao-yu ZHANG ; Wei-wei TANG ; Qing-yu HUANG
Journal of Forensic Medicine 2010;26(2):81-83
OBJECTIVE:
To examine the effects of cocaine on the activities of ATPase, LDH and SDH in cultured mouse splenocytes in vitro.
METHODS:
The ATPase, LDH and SDH activities in mouse splenocytes were detected at day 7 after continuous culturing the mouse cells exposed to cocaine hydrochloride in final concentration of 10, 20 and 100 microg/mL in vitro.
RESULTS:
The activities of ATPase, LDH and SDH in mouse splenocytes exposed to cocaine hydrochloride in final concentration of 10, 20 and 100 microg/mL were significantly decreased after continuous culturing for 7 days.
CONCLUSION
The present study demonstrated that cocaine could inhibit the activities of ATPase, LDH and SDH in cultured splenocytes in vitro.
Adenosine Triphosphatases/metabolism*
;
Animals
;
Cells, Cultured
;
Cocaine/pharmacology*
;
Dose-Response Relationship, Drug
;
L-Lactate Dehydrogenase/metabolism*
;
Male
;
Mice
;
Mice, Inbred Strains
;
Spleen/enzymology*
;
Succinate Dehydrogenase/metabolism*
7.Succinic semialdehyde dehydrogenase deficiency.
Xiao-Lu DENG ; Fei YIN ; Qiu-Lian XIANG ; Chen-Tao LIU ; Jing PENG
Chinese Journal of Contemporary Pediatrics 2011;13(9):740-742
Succinic semialdehyde dehydrogenase (SSADH) deficiency is a rare autosomal recessive disorder. This paper reports three cases of SSADH deficiency in infants. The infants developed the symptoms including developmental delay, intellectual disability, hypotonia, hyporeflexia and seizures. The electroencephalogram (EEG) showed background slowing and focal spike discharges in all of 3 patients. Head magnetic resonance imaging (MRI) demonstrated abnormalities in 2 patients, including basal ganglia damage and increased T2-weighted signal in bilateral cerebral peduncles. Urinary organic acid analysis with gas chromatography-mass spectrometry (GC-MS) revealed increased levels of 4-hydroxybutyrate (GHB) in 3 patients. SSADH deficiency was definitely diagnosed based on the clinical manifestations and the results of urinary organic acid analysis in the 3 children. It was concluded that early urine organic acid analysis is essential for children presenting with mental retardation, neuropsychiatric disturbance or epilepsy of unknown etiology.
Amino Acid Metabolism, Inborn Errors
;
diagnosis
;
therapy
;
Developmental Disabilities
;
Diagnosis, Differential
;
Female
;
Humans
;
Infant
;
Succinate-Semialdehyde Dehydrogenase
;
deficiency
8.The development of research on enzymes related to morphine-dependent.
Journal of Forensic Medicine 2005;21(2):152-158
The mechanism of morphine dependent is a complex Procedure. It involves in many complex mechanisms such as the ultra-structure of synapse of special brain areas, neurotransmitter, enzymology, and so on. These mechanisms have closely correlation. In this paper we reveiwed the development in enzymological mechanism of morphine dependent enzymes including protein kinase (PK), nitric oxide synthase (NOS), superoxide dismutase (SOD), adenylate cyclase (AC), Succinate dehydrogenase (SDH)and 3beta-Hydroxy steroid dehydrogenase (3beta-HSD).
Adenylyl Cyclases/metabolism*
;
Animals
;
Brain/enzymology*
;
Morphine Dependence/pathology*
;
Nitric Oxide Synthase/metabolism*
;
Protein Kinases/metabolism*
;
Substance Withdrawal Syndrome/metabolism*
;
Succinate Dehydrogenase/metabolism*
;
Superoxide Dismutase/metabolism*
;
Synapses/enzymology*
9.Effects of soybean isoflavones on the energy metabolism of swimming mice.
Bing-Nan DENG ; Jing-Ran SUN ; Hong JIN ; Hong-Jing NIE ; Rui-Feng DUAN ; Lie LIU ; Zhi-Xian GAO ; Zhao-Li CHEN
Chinese Journal of Applied Physiology 2018;34(1):39-42
OBJECTIVE:
To establish an animal model for loaded swimming, so as to investigate the energy metabolism effects of soybean isoflavones (SI) on swimming mice.
METHODS:
Thirty male Kunming mice were randomly divided into three groups:normal control, swimming group, and swimming+SI group. The normal control group mice were fed a basic AIN-93M diet, the SI groups were supplied with soybean isoflavones(4 g/kg).Two weeks later, the mice were forced to swim for an hour,and then all the mice were killed, the samples of blood, liver and muscles of hind were collected.The serum contents of lactic acid(Lac), the activities of lactic dehydrogenase (LDH), succinate dehydrogenase (SDH), creatine kinase (CK) and ATPase were measured.
RESULTS:
Compared with normal control,the serum content of Lac was significantly improved in the group of the swimming control and SI(<0.05),the activity of LDH in the serum was obviously improved in the group of the swimming control and SI, and the activity of CK and SDH were both significantly improved in the group of the swimming control and SI except the activity of SDH in the liver of the group SI; compared with the swimming control,the serum contents of Lac,the activities of LDH, ATPase, SDH, CK were obviously improved(<0.05).
CONCLUSIONS
Soybean isoflavones can improve the energy metabolism,antioxidant capacity of the swimming mice.
Adenosine Triphosphatases
;
blood
;
Animals
;
Creatine Kinase
;
blood
;
Energy Metabolism
;
Isoflavones
;
pharmacology
;
L-Lactate Dehydrogenase
;
blood
;
Lactic Acid
;
blood
;
Male
;
Mice
;
Random Allocation
;
Soybeans
;
chemistry
;
Succinate Dehydrogenase
;
blood
;
Swimming