1.A Randomized,Double-blind,Multi-center Clinical Trial of Tengfu Jiangya Tablets Combined with Valsartan/Amlodipine in Treatment of Grade 2 Hypertension with Hyperactivity of Liver Yang
Yuchen QI ; Yunlun LI ; Yujie WU ; Haiquan WANG ; Yu WANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(9):83-92
ObjectiveTo observe the clinical efficacy and safety of Tengfu Jiangya tablets combined with valsartan/amlodipine in the treatment of grade 2 hypertension with liver Yang hyperactivity syndrome. MethodAccording to a randomized,double-blind,and placebo parallel control method,288 patients with grade 2 hypertension with liver Yang hyperactivity from 7 subcenters were included. They were randomly divided into an observation group (144 cases) and a control group (144 cases),and then treated with Tengfu Jiangya tablets combined with valsartan/amlodipine and placebo combined with valsartan/amlodipine,respectively. The efficacy was evaluated after four weeks of intervention. During the experiment,the safety indicators and adverse reaction events of the subjects were recorded for safety evaluation,and the efficacy indicators and TCM syndrome scores were recorded for effectiveness evaluation. Sensitivity analysis was also conducted on the statistical results of the main efficacy indicators such as blood pressure (BP) compliance rate to ensure the accuracy of the analysis results. 88 groups of blood samples from each of the treatment and control groups were included as test subjects. Fasting blood samples were collected from the patients in the clinical trial on the day before and after medication,and enzyme linked immunosorbent assay (ELISA) was performed on the treated serum. The levels of arachidonic acid (AA),thromboxane B2 (TXB2),and prostaglandin E2 (PGE2) in the serum of the patients before and after treatment were measured to explore the regulation of inflammatory factors in the body by Tengfu Jiangya tablets. ResultA total of 271 patients (133 in the observation group and 138 in the control group) completed the trial. There was no statistically significant difference before and after treatment in such safety indicators as the blood routine (white blood cells,red blood cells,and platelets),urine routine (urinary protein and urinary red blood cells),alanine aminotransferase,aspartate aminotransferase,creatinine,urea,and abnormal electrocardiogram,and no serious adverse reactions were observed. After four weeks,the systolic blood pressure (SBP) difference and diastolic blood pressure (DBP) difference of patients in the observation group were greater than those in the control group(P<0.01). According to the criteria for determining the antihypertensive effect,the overall response rate in the observation group[89.47%(119/133)] was higher than that in the control group[57.97%(80/138)] (Z=2.593,P<0.01). The SBP compliance rate was 61.65%(82/133) and 37.68%(52/138) in the observation group and control group, respectively. The DBP compliance rate in the observation group was 78.20%(104/133),while in the control group it was 55.07%(76/138). The overall BP compliance rate in the observation group was 48.12%(64/133),while in the control group it was 23.19%(32/138). The BP compliance rates in the observation group were all significantly higher than those in the control group(χ2=15.571,16.236,18.404,P<0.01). According to the criteria for evaluating the therapeutic effect of TCM syndrome integration,the overall response rate of the observation group[57.89%(77/133)] was higher than that of the control group[38.41%(53/133)] (Z=-3.172,P<0.01).Compared with those before treatment, the levels of serum AA and TXB2 in the two groups were significantly decreased after treatment (P<0.01), and the level of PGE2 in the observation group was significantly increased (P<0.01). Compared with those of the control group after treatment, the levels of AA and TXB2 in the observation group were significantly decreased, while the level of PGE2 was significantly increased (P<0.01). The results suggest that Tengfu Jiangya tablets can effectively reduce inflammatory factors,reduce the production of inflammatory mediators,and thus prevent the occurrence of inflammatory reactions in the treatment of patients with grade 2 hypertension. ConclusionTengfu Jiangya tablets can more effectively reduce patients' SBP and DBP,improve their BP compliance rates,and improve their TCM syndromes in the treatment of grade 2 hypertension with liver Yang hyperactivity. Its clinical application is safe. Tengfu Jiangya tablets has outstanding clinical efficacy and can be used as an effective intervention method for the treatment of grade 2 hypertension with liver Yang hyperactivity syndrome.
2.Value of digital X-ray radiography and SPECT radionuclide bone imaging in diagnosing intertrochanteric fracture of femur
Xuewei HAO ; Fuming WU ; Jingyang SU ; Haiquan YU
China Medical Equipment 2024;21(1):51-54
Objective:To investigate the application value of digital X-ray radiotherapy and single photon emission computed tomography(SPECT)radionuclide bone imaging in the diagnosis of intertrochanteric fracture of femur.Methods:A total of 70 patients who were diagnosed as intertrochanteric fracture of femur in Shijiazhuang People's hospital from January 2017 to December 2020 were selected,and they were divided into study group and control group according to random number method,with 35 cases in each group.Digital X-ray radiography combined with SPECT radionuclide bone imaging was used in the study group,and digital X-ray radiography was used in the control group.And then,the accuracy of the diagnostic results between the two groups were compared.Results:The radiographic examination indicated that 31 cases(88.6%)of 35 cases in the study group were confirmed,and 24 cases(68.6%)of 35 cases in the control group were confirmed.The diagnostic accuracy rate of the study group was significantly higher than that of the control group,and the difference was statistically significant(x2=4.158,P<0.05).The preoperative imaging diagnosis indicated that diagnosed number of fracture blocks in the study group was(3.57±0.50),which was significantly higher than that(2.67±0.40)in the control group,and the difference of that between two groups was statistically significant(t=8.315,P<0.05).In the actual intraoperative examining for bone continuity,5 cases were continuity and 30 cases were non-continuity.In the bone stability,13 cases were stability and 22 cases were non-stability.The study group was closer to intraoperative bone continuity and bone stability,and the differences of them between two groups were statistically significant(x2=12.857,4.644,P<0.05),respectively.Conclusion:The combination of digital X-ray radiography and SPECT radionuclide bone imaging has higher diagnostic value for intertrochanteric fracture of femur,which can provide important reference for clinical diagnosis.
3.Based on Network Pharmacology and Experimental Verification to Explore the Mechanism of Yangxin Decoction Anti-atherosclerosis
Haiquan HUANGFU ; Huichun HUANG ; Xiaoming SHANG ; Zhichao HUANG ; Weiqian LIAO ; Hairui YU
World Science and Technology-Modernization of Traditional Chinese Medicine 2024;26(4):1070-1079
Objective Based on network pharmacology and in vivo experiments,to explore the mechanism of Yangxin Decoction in treating arterial atherosclerosis.Methods In the network pharmacology part,TCMSP database is used to screen the drug active ingredients and corresponding targets of Yangxin Decoction,Gene Cards,DisGeNet,OMIM,TTD database is used to screen atherosclerosis disease targets,the Evenn platform for interactive mapping to obtain drug disease intersection targets.Cytoscape 3.7.2 software is used to build a drug active ingredient core target disease interaction network diagram,The intersection target points are imported into STRING database to obtain PPI network diagram,and the Cytascape software is used for visualization processing.The metascape v3.5 platform is used for GO and KEGG enrichment analysis,and the micro-signal platform is used for visualization processing.In vivo experiment:ApoE-/-mice established atherosclerosis animal models through high-fat diet.The model mice were randomly divided into model group(Model),low-dose Yangxin Decoction group(YXT-L),and high-dose Yangxin Decoction group(YXT-H).C57BL/6 mice were taken as the control group,YXT-L group and YXT-H group were respectively given 2.6 g·kg-1·d-1 and 5.2 g·kg-1·d-1 of Yangxin Decoction extract aqueous solution,and the control group and model group were given equal volume distilled water for 4 weeks.Oil red O staining was used to observe the plaque area of aortic sinus,and ELISA was used to detect serum IL-6 and IL-1β,Caspase-3,VEGF levels,TG,TC,LDL-C,HDL-C levels of blood lipids detected by automatic biochemical instrument,and NF-κB p65,TNF-α,IKKβ Protein expression of aorta detected by Western blot.Results Network pharmacology:105 active ingredients of Yangxin Decoction were screened out,including 535 corresponding targets,4921 atherosclerotic disease targets,162 drug disease intersection targets,and the top 10 targets include AKT1,TNF,IL-6,VEGFA,IL-1β,TP53,JUN,CASP3,PPARG,PTGS2.A total of 2224 items were obtained from and GO analysis,including 1946 biological processes,100 cell components and 178 molecular functions.A total of 216 signal pathways were obtained by KEGG signal pathway enrichment analysis,mainly involving fluid shear stress,atherosclerosis,NF-κB signaling pathway,cAMP signaling pathway,diabetes cardiomyopathy,cysteine and methionine metabolism,VEGF signaling pathway,calcium signaling pathway,etc.In vitro experiment:Yangxin Decoction reduces serum TG,TC,LDL-C in ApoE-/-atherosclerosis mice in a dose-dependent manner,increases HDL-C level,reduces aortic sinus plaque area,and reduces serum IL-6,IL-1β,Caspase-3 and VEGF level;Inhibition of aortic NF-κB p65,TNF-α,IKKβ Protein expression.Conclusion Yangxin Decoction may inhibit TNF-α/IKKβ/NF-κB signaling pathway plays an anti-atherosclerosis role by regulating lipid metabolism,inhibiting inflammatory reaction,regulating cell apoptosis,etc.
4.Meta-analysis of the clinical efficacy and safety of anatomical reconstruction and repair in the treatment of chronic lateral instability of the ankle joint
Haiquan ZENG ; Weijin MIAO ; Ming YU ; Min WANG ; Wen WANG
China Modern Doctor 2024;62(30):13-20
Objective To evaluate the clinical efficacy and safety of anatomical repair and anatomical reconstruction through Meta-analysis.Methods Systematically search PubMed,Embase,Wanfang database,China National Knowledge Infrastructure and other databases were used to obtain the relevant literature in the treatment of chronic lateral instability of the ankle joint with anatomical repair and reconstruction from the establishment of each database to December 2022,and Revman 5.4 and R 4.2 software were used for Meta-analysis.Results Eight articles were finally included in this paper.All were randomized controlled trials.The results of Meta-analysis showed that:there were statistically significant differences between anatomical repair and anatomical reconstruction in the anterior talar translation (MD=0.54,95%CI:0.27-0.81,P<0.01),talus tilt angle (MD=0.36,95%CI:0.07-0.66,P=0.02),and American Orthopaedic Foot & Ankle Society scale (MD=-4.79,95%CI:-6.58--3.01,P<0.01),but there was no statistically significant difference in complications (RR=1.13,95%CI:0.32-3.97,P=0.85).Conclusion Anatomical reconstruction is superior to anatomical repair in clinical efficacy,while the risk of complications is comparable between two.
5.Production of limonene and its derivative in Saccharomyces cerevisiae via metabolic engineering.
Yao HUANG ; Haiquan YANG ; Wei SHEN ; Yuanyuan XIA ; Yu CAO ; Xianzhong CHEN
Chinese Journal of Biotechnology 2023;39(11):4647-4662
Limonene and its derivative perillic acid are widely used in food, cosmetics, health products, medicine and other industries as important bioactive natural products. However, inefficient plant extraction and high energy-consuming chemical synthesis hamper the industrial production of limonene and perillic acid. In this study, limonene synthase from Mentha spicata was expressed in Saccharomyces cerevisiae by peroxisome compartmentalization, and the yield of limonene was 0.038 mg/L. The genes involved in limonene synthesis, ERG10, ERG13, tHMGR, ERG12, ERG8, IDI1, MVD1, ERG20ww and tLS, were step-wise expressed via modular engineering to study their effects on limonene yield. The yield of limonene increased to 1.14 mg/L by increasing the precursor module. Using the plasmid with high copy number to express the above key genes, the yield of limonene significantly increased up to 86.74 mg/L, which was 4 337 times higher than that of the original strain. Using the limonene-producing strain as the starting strain, the production of perillic acid was successfully achieved by expressing cytochrome P450 enzyme gene from Salvia miltiorrhiza, and the yield reached 4.42 mg/L. The results may facilitate the construction of cell factory with high yield of monoterpene products by S. cerevisiae.
Saccharomyces cerevisiae/metabolism*
;
Limonene/metabolism*
;
Metabolic Engineering
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Monoterpenes/metabolism*
6.Clinical efficacy of laparoscopic sleeve gastrectomy in obesity patients of different grades
Na ZHANG ; Shengjun ZHANG ; Hongjuan MAI ; Qizhong CHEN ; Rui NI ; Yongtao YU ; Shensi CHEN ; Haiquan QIAN
Chinese Journal of Digestive Surgery 2023;22(8):987-995
Objective:To investigate the clinical efficacy of laparoscopic sleeve gastrectomy (LSG) in obesity patients of different grades.Methods:The retrospective and descriptive study was conducted. The clinical data of 139 obesity patients of different grades who were admitted to the General Hospital of Ningxia Medical University from January 2018 to December 2021 were collected. There were 37 males and 102 females, aged (32±9)years. Of the 139 patients, there were 45 cases of grade Ⅰ obesity, 44 cases of grade Ⅱ obesity and 50 cases of grade Ⅲ obesity, respectively. Obser-vation indicators: (1) intraoperative and postoperative situations in obesity patients of different grades; (2)hematological related indicators in obesity patients of different grades; (3)body quality related indicators in obesity patients of different grades. Measurement data with normal distribution were represented as Mean± SD, and comparison within three groups was conducted using the One-way Anova test and comparison between groups was conducted using the LSD test. Measurement data with skewed distribution were represented as M(range), and comparison within three groups was conducted using the Kruskal-Wallis H test. Count data were described as absolute numbers, and comparison between groups was performed using the chi-square test or Fisher exact probability. Repeated measurement data were analyzed by the repeated ANOVA. Results:(1) Intraoperative and postoperative situations in obesity patients of different grades. The operation time,volume of intraoperative blood loss,duration of postoperative hospital stay and cases readmitted within 30 days after surgery were 2.0(range, 1.5-2.0)hours,50(range, 50-100)mL, 5(range, 4-6)days and 2 in the 45 cases of grade Ⅰ obesity. The above indicators were 2.0(range, 1.5-2.0)hours, 60(range, 50-187)mL, 5(range, 4-6)days and 4 in the 44 cases of grade Ⅱ obesity and 2.0(range, 1.5-2.1)hours, 60(range, 50-135)mL, 5(range, 4-7)days and 4 in the 50 cases of grade Ⅲ obesity. There was no significant difference in the operation time, volume of intraoperative blood loss,duration of postoperative hospital stay among the three groups of patients ( H=4.11, 0.77, 3.59, P>0.05) and there was no significant difference in cases readmitted within 30 days after surgery among the three groups of patients ( P>0.05). (2) Hematological related indicators in obesity patients of different grades. All 139 patients were followed up at postoperative 3, 6 and 12 month. The total cholesterol (TC) were (4.5±0.9)mmol/L, (4.6±0.9)mmol/L, (4.3±0.8)mmol/L, (4.6±1.1)mmol/L at preoperative and postoperative 3, 6 and 12 month in cases of grade Ⅰ obesity. The above indicators were (4.5±0.8)mmol/L, (4.4±0.8)mmol/L, (4.4±1.0)mmol/L, (4.3±0.9)mmol/L in cases of grade Ⅱ obesity and (4.4±1.0)mmol/L, (4.7±1.1)mmol/L, (4.5±0.8)mmol/L, (4.4±0.5)mmol/L in cases of grade Ⅲ obesity. The above indicators among the three groups not meeting the mauchly′s test of sphericity ( χ2=20.81, P<0.05) and results of multi-variate test showed that there was no significant difference in the time effect, intergroup effect and interaction effect of TC among the three groups( Ftime=0.45, Fgroup=0.40, Finteraction=0.66, P>0.05). The triglyceride (TG) were (2.0±1.1)mmol/L, (1.3±0.4)mmol/L, (1.0±0.4)mmol/L, (1.0±0.4)mmol/L at preoperative and postoperative 3, 6 and 12 month in cases of grade Ⅰ obesity. The above indicators were (2.2±1.1)mmol/L, (1.5±0.5)mmol/L, (1.1±0.3)mmol/L, (1.0±0.3)mmol/L in cases of grade Ⅱ obesity and (2.3±1.1)mmol/L, (1.7±0.7)mmol/L, (1.4±0.6)mmol/L, (1.2±0.4)mmol/L in cases of grade Ⅲ obesity. The above indicators among the three groups not meeting the mauchly′s test of sphericity ( χ2=290.49, P<0.05) and results of multi-variate test showed that there were significant differences in the time effect, intergroup effect and interaction effect of TG among the three groups ( Ftime=80.44, Fgroup=4.13, Finteraction=2.67, P<0.05). The vitamin D were (12.9±5.9)μg/L, (16.5±5.9)μg/L, (18.0±6.3)μg/L, (20.1±5.7)μg/L at preoperative and postoperative 3, 6 and 12 month in cases of grade Ⅰ obesity. The above indicators were (11.5±4.4)μg/L, (17.1±5.0)μg/L, (18.2±5.6)μg/L, (20.2±6.6)μg/L in cases of grade Ⅱ obesity and (9.8±3.5)μg/L, (17.2±4.6)μg/L, (18.1±4.7)μg/L, (19.5±5.2)μg/L in cases of grade Ⅲ obesity. The above indicators among the three groups not meeting the mauchly′s test of sphericity ( χ2=53.07, P<0.05) and results of multi-variate test showed that there were significant differences in the time effect and interaction effect of vitamin D among the three groups ( Ftime=150.88, Finteraction=3.86, P<0.05)and there was no significant difference in the intergroup effect of vitamin D among the three groups ( Fgroup=0.35, P>0.05). (3) Body quality related indicators in obesity patients of different grades. The body mass and body mass index (BMI) were (88±8)kg, (71±8)kg, (65±8)kg, (61±7)kg, (32±2)kg/m 2, (26±2)kg/m 2, (24±2)kg/m 2, (22±2)kg/m 2 at preoperative and postoperative 3, 6 and 12 month in cases of grade Ⅰ obesity. The above indicators were (106±11)kg, (82±8)kg, (75±9)kg, (70±9)kg, (37±1)kg/m 2, (29±2)kg/m 2, (26±2)kg/m 2, (25±3)kg/m 2 in cases of grade Ⅱ obesity and (131±20)kg, (101±15)kg, (89±13)kg, (79±12)kg, (45±6)kg/m 2, (35±5)kg/m 2, (31±4)kg/m 2, (27±4)kg/m 2 in cases of grade Ⅲ obesity. The above indicators among the three groups not meeting the mauchly′s test of sphericity ( χ2=194.60, 179.52, P<0.05) and results of multi-variate test showed that there were significant differences in the time effect, intergroup effect and interaction effect of body mass and BMI among the three groups( Ftime=492.59, 543.86, Fgroup=89.13, 95.91, Finteraction=13.97, 13.32, P<0.05). The percen-tage of excess weight loss (EWL%) were 61%±16%,84%±21%,96%±23% at postoperative 3, 6 and 12 month in cases of grade Ⅰ obesity. The above indicators were 55%±7%,72%±16%,85%±19% in cases of grade Ⅱ obesity and 45%±12%,64%±15%,78%±7% in cases of grade Ⅲ obesity. The above indicators among the three groups not meeting the mauchly′s test of sphericity ( χ2=51.61, P<0.05) and results of multi-variate test showed that there were significant differences in the time effect and intergroup effect of EWL% among the three groups ( Ftime=374.52, Fgroup=15.69, P<0.05) and there was no significant difference in the interaction effect of EWL% among the three groups ( Finteraction=1.08, P>0.05). The percentage of total body weight loss (TWL%) were 19%±5%, 26%±6%, 30%±6% at postoperative 3, 6 and 12 month in cases of grade Ⅰ obesity. The above indicators were 21%±6%, 29%±6%, 34%±7% in cases of grade Ⅱ obesity and 22%±7%, 32%±7%, 39%±8% in cases of grade Ⅲ obesity. The above indicators among the three groups not meeting the mauchly′s test of sphericity ( χ2=58.54, P<0.05) and results of multi-variate test showed that there were significant differences in the time effect, intergroup effect and interaction effect of TWL% among the three groups ( Ftime=290.61, Fgroup=12.32, Finteraction=5.49, P<0.05). The waist to hip ratio (WHR) and visceral fat area (VFA) were 0.92±0.04, 0.86±0.03, 0.84±0.03, 0.83±0.03, (129±19)cm 2, (79±17)cm 2, (63±15)cm 2, (57±14)cm 2 at preoperative and postoperative 3, 6 and 12 month in cases of grade Ⅰ obesity. The above indi-cators were 0.98±0.05, 0.90±0.05, 0.87±0.05, 0.86±0.05, (169±20)cm 2, (105±23)cm 2, (85±20)cm 2, (73±20)cm 2 in cases of grade Ⅱ obesity and 1.05±0.09, 0.94±0.06, 0.91±0.06, 0.89±0.05, (218±42)cm 2, (144±35)cm 2, (114±26)cm 2, (96±19)cm 2 in cases of grade Ⅲ obesity. The above indicators among the three groups not meeting the mauchly′s test of sphericity ( χ2 =289.99, 191.92, P<0.05) and results of multi-variate test showed that there were significant differences in the time effect, intergroup effect and interaction effect of WHR and VFA among the three groups ( Ftime=361.39, 707.60, Fgroup=34.28, 12.69, Finteraction=8.31, 94.89, P<0.05). Conclusion:Treatment of obesity patients of different grades with LSG can improve patients′ TG and vitamin D levels, and reduce patients′ body mass, BMI, EWL%, TWL%, WHR and VFA.
7.Effect of hydrogen on lung injury induced by extremity ischemia-reperfusion in elderly patients
Tongxiang NIU ; Lin SU ; Haiquan HAO ; Chao QIN ; Keliang XIE ; Yonghao YU
Chinese Journal of Anesthesiology 2021;41(6):647-650
Objective:To evaluate the effect of hydrogen on lung injury induced by extremity ischemia-reperfusion (I/R) in elderly patients.Methods:Sixty American Society of Anesthesiologists physical status Ⅱ or Ⅲ elderly patients, aged 65-75 yr, with height 155-180 cm, weighing 50-75 kg, undergoing lower limb surgery under spinal anesthesia, were divided into 2 groups ( n=30 each) using a random number table method: hydrogen inhalation group (H group) and control group (C group). In H group, 67% hydrogen-33% oxygen was inhaled through the nasal catheter until the end of surgery starting from the completion of anesthesia.In group C, 33% oxygen was inhaled through the nasal catheter until the end of surgery after the completion of anesthesia.Blood samples from the radial artery were collected before anesthesia and at 60 min after tourniquet deflation.Blood gas analysis was performed to determine and record arterial oxygen partial pressure (PaO 2) and arterial carbon dioxide partial pressure (PaCO 2), and alveolar-arterial partial pressure of oxygen difference (A-aDO 2), oxygenation index (OI) and respiratory index (RI) were calculated.Pulmonary surfactant protein D (SP-D) and interleukin-6 (IL-6) concentrations in serum were measured by enzyme-linked immuno sorbent assay.ICU stay time and incidence of pulmonary complications within 7 days after operation were recorded. Results:Compared with group C, PaO 2 and OI were significantly increased, RI and A-aDO 2 were decreased, SP-D and IL-6 concentrations in serum were decreased at 60 min after tourniquet deflation, and ICU stay time was shortened ( P<0.05), and no significant change was found in the incidence of pulmonary complications within 7 days after surgery in group H ( P>0.05). Conclusion:Hydrogen can reduce the lung injury induced by extremity I/R, and the mechanism may be related to the reduction of inflammatory response in elderly patients.
8.Hemodynamic Analysis on Proximal End of the Aortic Dissection with Different Rupture Shapes
Yu XUE ; Qingsong HAN ; Yongzhi GONG ; Guizhen BAO ; Shijie GUO ; Haiquan FENG ; Xiaotian WANG ; Wei WEI
Journal of Medical Biomechanics 2021;36(5):E751-E756
Objective To explore hemodynamic performance of the aortic dissection after lesions, so as to provide a more scientific basis for patient treatment. Methods Based on computed tomography angiography (CTA) image data from a patient with complex Stanford B-type aortic dissection, the personalized aortic dissection models with different rupture shapes (H-type, O-type, and V-type) at proximal end of the aortic dissection were established. Combined with computational fluid dynamics (CFD) and morphological analysis method, distributions of the velocity at rupture section, the blood flow, the wall pressure and the wall shear stress (WSS) were analyzed. Results The flow velocity, the highest pressure difference and the WSS proportion at entrance of the H-shaped rupture showed larger hemodynamic parameters than those of the other two types. The risk of dissection rupture for type H was the largest, while type V was in the middle, and type O was the smallest. Conclusions This study provides an effective reference for further numerical analysis the cases and formulation of treatment plans.
9.Characterization of a sucrose phosphorylase from Leuconostoc mesenterides for the synthesis of α-arbutin.
Xiaoyu LI ; Yuanyuan XIA ; Wei SHEN ; Haiquan YANG ; Yu CAO ; Xianzhong CHEN
Chinese Journal of Biotechnology 2020;36(8):1546-1555
Sucrose phosphorylase (SPase) gene from Leuconostoc mesenteroides ATCC 12291 was synthesised after codon optimization, and inserted into pET-28a plasmid to generate pET-28a-spase. The recombinant strain Escherichia coli BL21 (DE3)/pET-28a-spase was induced for Spase expression. The recombinant protein Spase was purified and characterized. The specific enzyme activity of SPase was 213.98 U/mg, the purification ratio was 1.47-fold, and the enzyme activity recovery rate was 87.80%. The optimal temperature and the optimal pH of the SPase were identified to be 45 °C and 6.5 respectively, and Km, Vmax and kcat of the SPase for sucrose was 128.8 mmol/L, 2.167 μmol/(mL·min), and 39 237.86 min-1. The recombinant SPase was used for α-arbutin production from hydroquinone and the reaction process was evaluated. The optimal conditions for synthesis of α-arbutin by SPase were 40 g/L hydroquinone, 5:1 molar ratio of sucrose and hydroquinone, and 250 U/mL recombinant SPase at pH 7.0 and 30 °C for 24 h in the dark, and then 500 U/mL glucoamylase was added at 40°C for 2.5 h. Under the optimized process, the yield of α-arbutin reached 98 g/L, and the hydroquinone conversion rate was close to 99%. In summary, the recombinant SPase was cloned and characterized, and its application for α-arbutin production was feasible.

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