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1.
Microvasc Res ; 139: 104263, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34655603

RESUMO

Cannabinoids are reported to regulate cardiovascular functions. Cannabinoid receptors 1 (CB1Rs) are widely expressed in both the neuronal system and vascular system, but the contribution of CB1Rs in vascular smooth muscle (CB1RSM) to cardiovascular functions is not clear yet. In this research, we analyzed the effects of CB1RSM on blood pressure, vasoconstriction, and vasodilation abilities by using conditionally CB1R knockout mice (CB1RSMKO). The results show no significant difference in basal blood pressure between the conscious CB1RSMKO and control mice, indicating that CB1RSM is not essential for basal blood pressure maintenance. The constriction of the CB1RSMKO mesenteric artery in vitro was not significantly altered compared with that of the control mice. In contrast, the relaxation to CB1R agonist 2-AG or WIN55212-2 was decreased in CB1RSMKO vessels, suggesting that activation of CB1RSM mediates the vasodilation effect of cannabinoids. Ischemia stroke mouse model was used to further identify the potential function of CB1RSM in pathological conditions, and the results showed that the infarct volume in CB1RSMKO mice is significantly increased compared with the control littermates. These results suggest that vascular CB1R may not play a central role in basal vascular health maintenance but is protective in ischemia states, such as stroke. The protection function may be mediated, at least partly, by the relaxation effect of CB1RSM-dependent activities of endocannabinoids.

2.
Cell Rep ; 37(5): 109913, 2021 Nov 02.
Artigo em Inglês | MEDLINE | ID: mdl-34731618

RESUMO

Opiates produce a strong rewarding effect, but abstinence from opiate use emerges with severe negative emotions. Depression is one of the most frequent emotion disorders associated with opiate abstinence, which is thought to be a main cause for relapse. However, neurobiological bases of such an aversive emotion processing are poorly understood. Here, we find that morphine abstinence activates κ-opioid receptors (KORs) by increasing endogenous KOR ligand dynorphin expression in the amygdala, which in turn facilitates glutamate transporter 1 (GLT1) expression by activation of p38 mitogen-activated protein kinase (MAPK). Upregulation of GLT1 expression contributes to opiate-abstinence-elicited depressive-like behaviors through modulating amygdalar glutamatergic inputs to the nucleus accumbens (NAc). Intra-amygdala injection of GLT1 inhibitor DHK or knockdown of GLT1 expression in the amygdala significantly suppresses morphine-abstinence-induced depressive-like behaviors. Pharmacological and pharmacogenetic activation of amygdala-NAc projections prevents morphine-abstinence-induced behaviors. Overall, our study provides key molecular and circuit insights into the mechanisms of depression associated with opiate abstinence.

3.
World J Clin Cases ; 9(28): 8616-8626, 2021 Oct 06.
Artigo em Inglês | MEDLINE | ID: mdl-34754876

RESUMO

BACKGROUND: Primary intramedullary melanocytoma is an exceedingly rare type of primary melanocytic tumor in the central nervous system. Unfortunately, primary intramedullary melanocytoma lacks specificity in clinical symptoms and imaging features and there is currently no standard strategy for diagnosis or treatment. CASE SUMMARY: A 52-year-old male patient suffered from weakness and numbness involving the bilateral lower limbs for 18 mo, and defecation and erectile dysfunction for 6 mo. Furthermore, these symptoms started to worsen for the last 3 mo. Preoperative magnetic resonance imaging (MRI) revealed an intramedullary tumor located at the T9-T10 level. In subsequently surgery, the maximal safe resection extent approached to 98%. The lesion was confirmed to be melanocytoma by pathological examination. In addition, the possibility of original melanocytoma outside the spinal cord was excluded after the examination of the whole body. Therefore, a diagnosis of primary intramedullary melanocytoma was established. The patient refused to accept radiotherapy or Gamma Knife, but MRI examination on July 28, 2020 showed no sign of development. In addition, on April 10, 2021, the recent review showed that the disorder of defecation and lower limbs improved further but erectile dysfunction benefited a little from the surgery. CONCLUSION: After diagnosing intramedullary melanocytoma by postoperative pathology, the inspection of the whole body contributed to excluding the possibility of metastasis from other regions and further suggested a diagnosis of primary intramedullary melanocytoma. Complete resection, adjuvant radiation, and regular review are critical. In addition, maximal safe resection also benefits prognosis while the tumor is difficult to be resected totally.

4.
J Biotechnol ; 343: 7-14, 2021 Nov 09.
Artigo em Inglês | MEDLINE | ID: mdl-34763007

RESUMO

Transaminases catalyze the transfer of an amino group from a donor to a keto group of an acceptor substrate and are applicable to the asymmetric synthesis of herbicide L-phosphinothricin (L-PPT). Here, the important residue sites (C390, I22, V52, R141, Y138 and D239) of transaminase from Salmonella enterica (SeTA) were modified at the adjacency of the substrate-binding pocket to improve the enzyme activity. Among the constructed mutant library, the SeTA-Y138F mutant displayed higher activity than the wild-type enzyme. Compared to the wild-type, SeTA-Y138F showed improved catalytic efficiency with a 4.36-fold increase. The Km and kcat of SeTA -Y138F toward 4-(hydroxy(methyl) phosphoryl)-2-oxobutanoic acid (PPO) were 26.39 mM and 34.28 s-1, respectively. Subsequently, the three-enzyme co-expression system of E. coli BL21 (DE3)/pACYCDuet-SeTA-Y138F/pETDuet-AlaDH-BsGDH was developed by combining a alanine dehydrogenase (AlaDH) to recycle the byproduct of amino donor, a glucose dehydrogenase (BsGDH) for cofactor recycling. Under the optimized conditions, an excellent L-PPT yield of 90.8% was achieved by the whole-cell biotransformation with 500 mM PPO. It exhibited the tri-enzymatic coupling system was potential for effective production of target L-PPT.

5.
Front Cardiovasc Med ; 8: 751182, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34805305

RESUMO

Background: Studies have shown inconsistent associations between serum uric acid (SUA) levels and mortality in peritoneal dialysis (PD) patients. We conducted this meta-analysis to determine whether SUA levels were associated with cardiovascular or all-cause mortality in PD patients. Methods: PubMed, Embase, Web of Science, the Cochrane Library, CNKI, VIP, Wanfang Database, and trial registry databases were systematically searched up to April 11, 2021. Cohort studies of SUA levels and cardiovascular or all-cause mortality in PD patients were obtained. Random effect models were used to calculate the pooled adjusted hazard ratio (HR) and corresponding 95% confidence interval (CI). Sensitivity analyses were conducted to assess the robustness of the pooled results. Subgroup analyses and meta-regression analyses were performed to explore the sources of heterogeneity. Funnel plots, Begg's tests, and Egger's tests were conducted to evaluate potential publication bias. The GRADE approach was used to rate the certainty of evidence. This study was registered with PROSPERO, CRD42021268739. Results: Seven studies covering 18,113 PD patients were included. Compared with the middle SUA levels, high SUA levels increased the risk of all-cause mortality (HR = 1.74, 95%CI: 1.26-2.40, I 2 = 34.8%, τ2 = 0.03), low SUA levels were not statistically significant with the risk of all-cause or cardiovascular mortality (HR = 1.04, 95%CI: 0.84-1.29, I 2 = 43.8%, τ2 = 0.03; HR = 0.89, 95%CI: 0.65-1.23, I 2 = 36.3%, τ2 = 0.04; respectively). Compared with the low SUA levels, high SUA levels were not statistically associated with an increased risk of all-cause or cardiovascular mortality (HR = 1.19, 95%CI: 0.59-2.40, I 2 = 88.2%, τ2 = 0.44; HR = 1.22, 95%CI: 0.39-3.85, I 2 = 89.3%, τ2 = 0.92; respectively). Conclusion: Compared with middle SUA levels, high SUA levels are associated with an increased risk of all-cause mortality in PD patients. SUA levels may not be associated with cardiovascular mortality. More high-level studies, especially randomized controlled trials, are needed to determine the association between SUA levels and cardiovascular or all-cause mortality in PD patients. Systematic Review Registration: https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42021268739, identifier: CRD42021268739.

6.
Biotechnol Prog ; : e3218, 2021 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-34601810

RESUMO

The Candida antarctica lipase B (CALB) was embedded in the metal-organic framework, zeolitic imidazolate framework-8 (ZIF-8), and applied in the enzymatic synthesis of L-ascorbic acid palmitate (ASP) for the first time. The obtained CALB@ZIF-8 achieved the enzyme loading of 80 mg g-1 with 11.3 U g-1 (dry weight) unit activity, 59.8% activity recovery, and 92.7% immobilization yield. Under the optimal condition, ASP was synthesized with over 75.9% conversion of L-ascorbic acid in a 10-batch reaction. Continuous synthesis of ASP was subsequently performed in a packed bed bioreactor with an outstanding average space-time yield of 58.1 g L-1  h-1 , which was higher than ever reported continuous ASP biosynthesis reactions.

7.
Biotechnol J ; : e2100431, 2021 Oct 27.
Artigo em Inglês | MEDLINE | ID: mdl-34705325

RESUMO

BACKGROUND: d-Pantothenate (DPA) is an important functional chemical that has been widely applied in healthcare, cosmetics, animal food, and feed industries. METHODS AND RESULTS: In this study, a high-yield DPA-producing strain was constructed by metabolic engineering strategies with targeting metabolic driving and by-products minimization. The metabolic driving force of push and pull was firstly obtained to improve the production of DPA via enrichment of precursor pool and synthetic pathway, accumulating 4.29 g L-1 DPA in shake flask fermentation. To eliminate the metabolic pressure on DPA production, an amino throttling system was proposed and successfully attenuated the synthesis of four competitive amino acids by a single-step regulation of gdhA. Further minimization of acetate was carried out by pta deletion, and utilization of ß-alanine was improved via enhancing its uptake system with producing 5.78 g L-1 DPA. Finally, the engineered strain produced 66.39 g L-1 DPA with ß-alanine addition in fermentor under fed-batch fermentation. CONCLUSION: This study paved a foundation for the industrial production of DPA.

8.
Enzyme Microb Technol ; 152: 109931, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34688091

RESUMO

Glucose isomerase (GI) is a key enzyme in the preparation of high fructose corn syrup (HFCS). In this study, a mutant TEGI-M-L38 M/V137 L (TEGI-M2) of glucose isomerase (TEGI-M) originated from Thermoanaerobacter ethanalicus CCSD1 was obtained by site-directed mutagenesis. The TEGI-M2 showed an optimal activity at 85 ℃ and pH 6.5 with the divalent cations Co2+ and Mg2+. The structural differences between TEGI-M and TEGI-M2 were investigated based on the homology modeling and molecular docking, to elucidate the mechanism of improvement in the enzymatic properties. Compared with the original enzyme, the TEGI-M2 showed a 2.0-fold increased enzyme activity and a decreased Km from 234.2 mM to 85.9 mM. Finally, the application of mutant TEGI-M2 in HFCS one-step biosynthesis was attempted, resulting in a d-fructose yield of 67.3 %, which was 14.3 % higher than that of TEGI-M. This improved catalytic performance of TEGI-M2 was of great importance for the industrial preparation of d-fructose in one-step process.


Assuntos
Aldose-Cetose Isomerases , Thermoanaerobacter , Aldose-Cetose Isomerases/genética , Simulação de Acoplamento Molecular , Mutagênese Sítio-Dirigida , Thermoanaerobacter/genética
9.
Bioresour Technol ; 342: 126050, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34597803

RESUMO

ß-Alanine is the only naturally occurring ß-amino acid, widely used in the fine chemical and pharmaceutical fields. In this study, metabolic design strategies were attempted in Escherichia coli W3110 for enhancing ß-alanine biosynthesis. Specifically, heterologous L-aspartate-α-decarboxylase was used, the aspartate kinase I and III involved in competitive pathways were down-regulated, the ß-alanine uptake system was disrupted, the phosphoenolpyruvate carboxylase was overexpressed, and the isocitrate lyase repressor repressing glyoxylate cycle shunt was delete, the glucose uptake system was modified, and the regeneration of amino donor was up-regulated. On this basis, a plasmid harboring the heterologous panD and aspB was constructed. The resultant strain ALA17/pTrc99a-panDBS-aspBCG could yield 4.20 g/L ß-alanine in shake flask and 43.94 g/L ß-alanine (a yield of 0.20 g/g glucose) in 5-L bioreactor via fed-batch cultivation. These modification strategies were proved effective and the constructed ß-alanine producer was a promising microbial cell factory for industrial production of ß-alanine.

10.
Artigo em Inglês | MEDLINE | ID: mdl-34519920

RESUMO

Sucrose isomerase (SIase) is a key enzyme used for the production of isomaltulose from sucrose. In this study, an SIase gene from Erwinia sp. Ejp617 (ErSIase) was heterologously expressed in Escherichia coli BL21(DE3), and the recombinant ErSIase was served as biocatalyst combined with the graphene oxide (GO) as carrier for ErSIase immobilization. The Fourier transform infrared spectroscopy, transmission electron microscope, and confocal laser microscopy analyses showed that ErSIase was successfully immobilized on the surface of GO to form ErSIase-GO. The loading capacity of ErSIase on GO reached up to 460 mg/g with a specific activity of 727.04 U/mg protein when the optimal immobilization time of 12 h and the ErSIase/GO ratio of 7.4:4 (w/w) were applied. A high conversion rate of 95.3% was reached from sucrose to isomaltulose using ErSIase-GO as biocatalyst with 600 g/L sucrose as substrate, after 180 min at 40 °C and pH 6.0. Moreover, stabilities of the immobilized ErSIase-GO in the aspects of thermal, pH, and storage were improved, and its activity after 10 batches still remained around 80% under the optimal conditions. The Km value of ErSIase-GO was 29.32 mM, and the kcat/Km was increased to 27.34 s-1 mM-1 when 0.1% (w/v) detergent NP40 was added. These results indicated that the ErSIase was well immobilized onto GO, and the ErSIase-GO is a promising biocatalyst with high operational stability and catalytic activity for industrial production of isomaltulose.

11.
J Agric Food Chem ; 69(40): 12002-12011, 2021 Oct 13.
Artigo em Inglês | MEDLINE | ID: mdl-34590865

RESUMO

Xylitol is a widely used natural sweetener for the reduction of excessive sugar consumption. However, concerns of xylitol consumption existed as it is a highly permeable substance in the colon that could cause diarrhea and other adverse symptoms. To assess the relationship between xylitol dosage and diarrhea, especially the influences of diarrhea on physiological characteristics, the immune system, and gut microbiota in rats, the control, low-dose (L), medium-dose (M), and high-dose (H) groups were fed with 0, 1, 3, and 10% of xylitol, respectively, correspondingly for 15 days, followed by a 7-day recovery. Only medium- and high-dose xylitol would cause diarrhea in rats. Quantitative imaging of colonic tissue and the expression levels of proinflammatory factors revealed a higher degree of immune responses in the rats from H groups but statistically stable in M groups, despite that light diarrhea was observed. A shift of the gut microbiota composition was observed in the rats from H groups, including significant decreases of genera Ruminococcaceae and Prevotella and a notable increase and colonization of Bacteroides, accompanied with changes of short-chain fatty acid production. Tolerance and adaptation to xylitol consumption were observed in a dose-dependent manner. Our findings demonstrate that diarrhea caused by the high dosage of xylitol can exert distinctive changes on gut microbiota and lay the foundation to explore the mechanism underlying the shift in gut microbiota composition.


Assuntos
Microbioma Gastrointestinal , Animais , Diarreia , Ácidos Graxos Voláteis , Intestinos , Ratos , Xilitol
12.
Int J Biol Macromol ; 187: 850-857, 2021 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-34339787

RESUMO

Echinocandin B deacylase (ECBD) from Actinoplanes utahensis can be applied to produce echinocandin B nucleus (ECBN), an essential intermediate of the echinocandins antifungal drugs such as anidulafungin. To date, the expression of ECBD has been limited to Streptomyces. To achieve the active expression of ECBD in Escherichia coli (E. coli), we constructed a plasmid carrying two subunits of ECBD for T7 RNA polymerase driven transcription of dicistron messenger after codon optimization. Subsequently, the introduction of peptide tags in the recombinant ECBD was adopted to reduce the formation of inclusion bodies and enhance the ECBD solubility. The peptide tags with the opposite electrostatic charge, hexa-lysine (6K) and GEGEG (GE), exhibited the best positive effect, which was verified by activity assay and structural simulation. After that, optimization of culture conditions and characterization of ECBD were conducted, the optimal pH and temperature were 7.0 and 60 °C. It is the first report concerning the functional expression of ECBD in the host E. coli. Our results reported here can provide a reference for the high-level expression of other deacylases with respect to a possible industrial application.

13.
3 Biotech ; 11(8): 387, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34350092

RESUMO

Microbial fermentation for enzyme production and then whole-cell catalysis for l-2-aminobutyric acid (l-ABA) production have huge potential for industrial application, but the catalytic capacities of cells are directly related to the fermentation process. Using a 50 L fermenter, the effects of initial glycerol concentration in the medium and rotating speed on cell catalytic capacity were investigated. Fermentation cells showed the best catalytic activity when the initial glycerol concentration was 12 g/L and the rotating speed was 250 rpm. Furthermore, we studied the difference between glycerol and glycerol mixtures as fed-batch media in pH-stat fed-batch fermentation. Results showed that glycerol had better catalytic activity than the glycerol mixture, and the effect of fed-batch fermentation was better than batch fermentation. Meanwhile, the enzyme activities of leucine dehydrogenase and formate dehydrogenase reached 129.87 U/g DCW and 437.02 U/g DCW, respectively, and the intracellular NAD(H) concentration reached 14.94 µmol/g DCW. Using the optimized fermentation parameters, amplified fermentation was then carried out in a 5000 L fermenter to demonstrate the industrial production of l-ABA by Escherichia coli BL21.

14.
Biotechnol J ; 16(10): e2100103, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34363653

RESUMO

BACKGROUND: Nitrile hydratase (NHase), was an excellent biocatalyst for the synthesis of amide compounds. NHase was typical heterodimeric metalloprotein, required of the assistance of activator for active expressions. In this work, we found a special Co-NHase HBA from Caldalkalibacillus thermarum, which had the ability of post-translational self-modification and could incorporate Co2+ into the catalytic center in the absence of activator. METHOD AND RESULTS: We simulated the movement of Co2+ in silico and established a hypothetical model to predict the Co2+ incorporation efficiency (XCo ) of NHases. According to the simulation results, NHase mutants with different positive charge distribution were constructed. Compared with wild-type, the Co2+ incorporation efficiency of K1 (M10K) was increased by 2.1-fold from 0.36 to 0.76, and the specific activity was increased by 3.2-fold from 136.3 to 432.0 U mg-1 , while mutant K1H1 (M10K, D11H) and K2H2 (M10K, D11H, E20K, N21H) lost the ability of post-translation self-modification. CONCLUSIONS AND IMPLICATIONS: The interactions of positively charged residues near the catalytic center, such as lysine with strong electrostatic repulsive interaction, arginine with weak electrostatic repulsive interaction and histidine with metal affinity, could limit the free diffusion of Co2+ in NHase and affect the efficiency of post-translational self-modification. This work also provided an effective strategy for protein engineering of NHases and other metalloenzymes.


Assuntos
Cobalto , Hidroliases , Bacillaceae , Cobalto/metabolismo , Hidroliases/genética , Processamento de Proteína Pós-Traducional
15.
Appl Biochem Biotechnol ; 193(11): 3624-3640, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34347249

RESUMO

ω-Transaminase (ω-TA) is an attractive biocatalyst for stereospecific preparation of amino acids and derivatives, but low catalytic efficiency and unfavorable substrate specificity hamper their industrial application. In this work, to obtain applicable (R)-ω-TA responsible for amination of α-keto acids substrates, the reactivities of eight previously synthesized ω-TAs toward pyruvate using (R)-α-methylbenzylamine ((R)-α-MBA) as amine donor were investigated, and Gibberella zeae TA (GzTA) with the highest (R)-TA activity and stereoselectivity was selected as starting scaffold for engineering. Site-directed mutagenesis around enzymatic active pocket and access tunnel identified three positive mutation sites, S214A, F113L, and V60A. Kinetic analysis synchronously with molecular docking revealed that these mutations afforded desirable alleviation of steric hindrance for pyruvate and α-MBA. Furthermore, the constructed single-, double-, and triple-mutant exhibited varying degrees of improved specificities toward bulkier α-keto acids. Using 2-oxo-2-phenylacetic acid (1d) as substrate, the conversion rate of triple-mutant F113L/V60A/S214A increased by 3.8-fold relative to that of wide-type GzTA. This study provided a practical engineering strategy for improving catalytic efficiency and substrate specificity of (R)-ω-TA. The obtained experience shed light on creating more industrial ω-TAs mutants that can accommodate structurally diverse substrates.

16.
J Biosci Bioeng ; 132(5): 451-459, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34420895

RESUMO

The biosynthetic pathway of l-methionine in microorganisms was complex and regulated at multiple levels. In this study, a two-step method for l-methionine production combined fermentation and biocatalysis was realized in one pot. The O-succinyl-l-homoserine (OSH) producing strain Escherichia coli W3110(DE3) ΔIJB∗TrcmetL/pTrc-metAfbr-Trc-thrAfbr-yjeH (ΔIJB) was constructed initially. OSH in the fermentation supernatant was then converted to l-methionine in the presence of O-succinyl-l-homoserine sulfhydrylase (OSHS) and sodium methanethiol. The titer of l-methionine could reach 21.1 g/L after 88 h (84 h fermentation and 4 h catalysis) in a two-step method (process 1). In a one-pot two-strain system (process 2), two strains ΔIJB and E. coli BL21(DE3)/pET28b-OSHS-cutinase were co-cultured, and 8.24 g/L l-methionine was obtained. In another one-pot one-strain system (process 3), strain E. coli ΔIJB/pET28b-OSHS-cutinase could co-express OSHS and cutinase during ΔIJB fermentation at the same time, obtaining 13.6 g/L l-methionine in a 5 L fermentor after 84 h. By comparing the three processes for l-methionine production based on the process 1, the simplified process in process 3 provided in this study showed potent in the large-scale production of l-methionine with convenient handling and production efficiency, but further works still need to be carried out to improve the l-methionine production.


Assuntos
Escherichia coli , Metionina , Catálise , Escherichia coli/metabolismo , Fermentação , Homosserina/análogos & derivados , Metionina/metabolismo
17.
Artigo em Inglês | MEDLINE | ID: mdl-34252982

RESUMO

Streptomyces nodosus is known as the main manufacturer of amphotericin B (AmB), which is an effective antifungal drug. It is verified that the optimization of fermentation conditions and key growth factors have a great impact on the yield of AmB. The AmB production of S. nodosus in cotton-seed meal (CM) medium was 1.6 times than that of beef-paste medium. The transcriptome analysis was used to analyze the effects of different nitrogen media and calcium on S. nodosus. Related genes of the EMP and TCA pathways, such as phosphofructokinase, pyruvate dehydrogenase, and citrate synthase, were upregulated in CM medium. The expression level of the PKS modules of the amphotericin synthesis gene cluster in beef-paste medium was higher. Other functional genes, such as amphGH and amphRIV, have the advantage of expressing in CM medium. Ca2+ promoted the upregulation of genes in metabolic pathways such as EMP pathway (pyruvate dehydrogenase), TCA pathway (citrate synthase), and amphotericin synthesis genes (PKS modules). The expression of WhiB family genes SNOD_RS 13310 and SNOD_RS 17625 was positively correlated with Ca2+ concentration. In addition, in the presence of calcium, the expression level of Sec transport system genes of S. nodosus was lower.

18.
3 Biotech ; 11(6): 299, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-34194892

RESUMO

Amphotericin, as an important macrolide antibiotic, is synthesized by Streptomyces nodosus. A high-yield S. nodosus ZJB2016050 was obtained by mutagenesis in our lab with the advantages of high yield, short fermentation cycle and few by-products, which was more suitable for industrial production. The fermentation differences in 50-tons bioreactor between S. nodosus ATCC14899 and S. nodosus ZJB2016050 were compared. The amphotericin B (AmB) yield of S. nodosus ZJB2016050 was 9.73 mg/g at 96 h, which was 30% higher than that of S. nodosus ATCC14899. The by-product amphotericin A (AmA) production of S. nodosus ZJB2016050 was 78% lower than that of S. nodosus ATCC14899. By performing whole-genome sequencing of S. nodosus ZJB2016050 and comparative genome analysis with the wild-type S. nodosus ATCC14899, it was found that the two strains have high synteny, but each has a special gene fragment. The genes functions of fragment were identified in the amino acid transport and metabolism, carbohydrate metabolism and lipid transport and metabolism. The gene functions of SNP (single nucleotide polymorphism) genes were identified in amino acid transport and metabolism, carbohydrate metabolism, coenzyme metabolism and secondary metabolites biosynthesis. The difference in signal-regulation and transcription may be the main reason for the differences between these two strains. Three GntR family egulatory factors of S. nodosus ATCC14899 may reduce the synthesis of amphotericin. Based on the analysis of comparative genomes, the effects of corn oil in S. nodosus ATCC14899 and S. nodosus ZJB2016050 were also compared. The results showed that corn oil can promote the fermentation of S. nodosus ZJB2016050. The S. nodosus ZJB2016050 may degrade fatty acids faster, and the degraded acyl-coenzyme can be used to synthesize amphotericin. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-021-02844-2.

19.
Bioprocess Biosyst Eng ; 44(11): 2387-2398, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34268619

RESUMO

Cordycepin is a major bioactive compound found in Cordyceps sinensis that exhibits a broad spectrum of biological activities. Here a Paecilomyces hepiali OR-1 strain was initially isolated from plateau soil for the bioproduction of cordycepin. Subsequently, strain modification including 60Co γ-ray and ultraviolet irradiation were employed to increase the cordycepin titer, resulted in a high-yield mutant strain P. hepiali ZJB18001 with the cordycepin content of 0.61 mg/gDCW, showing a 2.3-fold to that from the wild strain (0.26 mg/gDCW). Furthermore, medium screening based on Box-Behnken design and the response surface methodology facilitated the enhancement of cordycepin yield to the value of 0.96 mg/gDCW at 25 °C for 5 days in submerged cultivation with an optimized medium composition. The high cordycepin yield, rapid growth rate and stable genetic characteristics of P. hepiali ZJB18001 are beneficial in terms of costs and time for the industrialization of cordycepin production.

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