scholarly journals Engineering of Phenylacetaldehyde Reductase for Efficient Substrate Conversion in Concentrated 2-Propanol

2005 ◽  
Vol 71 (8) ◽  
pp. 4713-4720 ◽  
Author(s):  
Yoshihide Makino ◽  
Kousuke Inoue ◽  
Tohru Dairi ◽  
Nobuya Itoh

ABSTRACT Phenylacetaldehyde reductase (PAR) is suitable for the conversion of various aryl ketones and 2-alkanones to corresponding chiral alcohols. 2-Propanol acts as a substrate solvent and hydrogen donor of coupled cofactor regeneration during the conversion of substrates catalyzed by PAR. To improve the conversion efficiency in high concentrations of substrate and 2-propanol, selection of a PAR mutant library and the subsequent rearrangement of mutations were attempted. With only a single selection round and following the manual combination of advantageous mutations, PAR was successfully adapted for the conversion of high concentrations of substrate with concentrated 2-propanol. This method will be widely applicable for the engineering of enzymes potentially valuable for industry.

2020 ◽  
Vol 65 (9-10) ◽  
pp. 3-7
Author(s):  
V. V. Gostev ◽  
Yu. V. Sopova ◽  
O. S. Kalinogorskaya ◽  
M. E. Velizhanina ◽  
I. V. Lazareva ◽  
...  

Glycopeptides are the basis of the treatment of infections caused by MRSA (Methicillin-Resistant Staphylococcus aureus). Previously, it was demonstrated that antibiotic tolerant phenotypes are formed during selection of resistance under the influence of high concentrations of antibiotics. The present study uses a similar in vitro selection model with vancomycin. Clinical isolates of MRSA belonging to genetic lines ST8 and ST239, as well as the MSSA (ATCC29213) strain, were included in the experiment. Test isolates were incubated for five hours in a medium with a high concentration of vancomycin (50 μg/ml). Test cultures were grown on the medium without antibiotic for 18 hours after each exposure. A total of ten exposure cycles were performed. Vancomycin was characterized by bacteriostatic action; the proportion of surviving cells after exposure was 70–100%. After selection, there was a slight increase in the MIC to vancomycin (MIC 2 μg/ml), teicoplanin (MIC 1.5–3 μg/ml) and daptomycin (MIC 0.25–2 μg/ml). According to the results of PAP analysis, all strains showed an increase in the area under curve depending on the concentration of vancomycin after selection, while a heteroresistant phenotype (with PAP/AUC 0.9) was detected in three isolates. All isolates showed walK mutations (T188S, D235N, E261V, V380I, and G223D). Exposure to short-term shock concentrations of vancomycin promotes the formation of heteroresistance in both MRSA and MSSA. Formation of VISA phenotypes is possible during therapy with vancomycin.


2018 ◽  
pp. 23-30
Author(s):  
O.V. Gorbunova ◽  
◽  
N.P. Goncharuk ◽  
H.V. Zarichanska ◽  
N.A. Ermolovich ◽  
...  

Pregnancy against the background of bacterial vaginosis is accompanied by a high risk of obstetric and perinatal complications. Therefore, bacterial vaginosis must be treated in pregnant women. Screening and therapy are performed at the beginning of the II or III trimester of pregnancy. The survival of colonies of pathogenic microorganisms in biofilms is significantly increased, so they can remain viable even at high concentrations of antiseptic. The main advantage of using local combined antiseptic agents is the ability to achieve the maximum concentration of the antibiotic exactly in the place of the greatest accumulation of pathogens with the ability to influence biofilms. The objective: was to compare the efficacy and safety of various regimens of therapy with topical combined drugs (Lynda and Meratin Combi) in pregnant women with bacterial vaginosis. Materials and methods. The biocenosis of the vagina was investigated in 351 pregnant women in the II trimester. The diagnosis of bacterial vaginosis was established if the patient had any three of the Amsel criteria. Results. Microbiological screening of the vaginal biocenosis in the II trimester of pregnancy showed that normocenosis among the surveyed was 18.5%; bacterial vaginosis – 31.6%; vulvovaginal candidiasis – 26.5%; aerobic vaginitis – 22.8%, trichomonas vaginitis – 0.6%. Against the background of bacterial vaginosis, the threat of miscarriage, placental dysfunction occurred 6 times more often, anemia and preeclampsia three times more often, gestational pyelonephritis twice more often than in healthy pregnant women. In most patients, the sensitivity of the vaginal microflora to metronidazole and ornidazole is the same, but depends on the dose of the antiseptic, the sensitivity to miconazole was almost twice as high as to nystatin. This confirms the need for a differentiated selection of antiseptics for local therapy of bacterial vaginosis during pregnancy. Conclusion. A more rapid dynamics of the disappearance of the main symptoms of bacterial vaginosis and the normalization of the pH of the vaginal secretion were noted after the use of the drug Limenda. In order to prevent relapse of the disease, it is necessary to carry out the second stage of treatment with probiotics to restore its own lactoflora. Keywords: screening of vaginal biocenosis in the II trimester of pregnancy; complications of pregnancy against the background of bacterial vaginosis; treating bacterial vaginosis during pregnancy; biofilms; sensitivity of the vaginal microflora to antiseptics; topical treatment of bacterial vaginosis; Limenda; Meratin Kombi.


2012 ◽  
Vol 30 (4) ◽  
pp. 683-696 ◽  
Author(s):  
F.P. Guimaraes ◽  
R. Aguiar ◽  
J.A. Oliveira ◽  
J.A.A. Silva ◽  
D. Karam

The potential of three aquatic macrophytes, Azoll caroliniana, Salvinia minima and Lemna gibba, was evaluated in this work aimed at selection of plants to be used in remediation of environments contaminated by arsenic (As). The experiments were carried out in a greenhouse during six days in pots containing Hoagland solution (¼ ionic strength) at As concentrations of 0.5; 2.5 and 5.0 mg L-1. The three species showed greater As accumulation as the concentration of the metalloid in solution increased. However, a reduction was detected in fresh and dry mass gain when the plants were exposed to high As concentrations. The macrophytes showed differences in efficiency of removal of As in solution. A. caroliniana, S. minima and L. gibba accumulated, on average, 0.130; 0.200; and 1.397 mg mDM-1, respectively, when exposed to 5.0 mg L-1 of As. The macrophytes absorbed a greater quantity of As in solution with low phosphate content. The greater As concentration in L. gibba tissues lowered the chlorophyll and carotenoid contents as shown by the high chlorosis incidence. Lemna gibba also exhibited a decrease in leaf size, with the total chlorophyll and carotenoid synthesis not being affected by As in A. caroliniana. This species exhibited purplish leaves with high concentration of anthocyanin, whose presence suggested association to phosphate deficiency. Marginal necrosis occurred on S. minima floating leaves, with the released daughter-plants not showing any visual symptoms during the treatment. The percentage of As removed from the solution decreased when the plants were exposed to high concentrations of the pollutant. Among the three species studied, only L. gibba could be considered an As hyper-accumulator. The use of this plant species for remediation of aquatic environments was shown to be limited and requires further investigation.


2021 ◽  
Vol 68 (1) ◽  
Author(s):  
R. Jannathulla ◽  
O. Sravanthi ◽  
H. Imran Khan ◽  
H. Syed Moomeen ◽  
A. Gomathi ◽  
...  

Aquaculture serves as a sustainable source of good quality wholesome food and major input is feed. In recent years, considerable quantities of plant sources have been attempted for inclusion in shrimp feed, due to high demand and high cost of fishmeal. Plant-based ingredients are in general poor in attractability and palatability to aquatic species compared to marine sources. It is desirable to develop an economical and nutritious feed that is attractive and palatable for ensuring reduction of feed wastage. Chemoattractants nowadays are unavoidable in commercial shrimp feeds and are included either individually or in combination. The present review revealed that the various marine sources, in particular krill meal would be more effective for aquatic species, compared to those derived from vegetable origin and chemical compounds. In addition to acting as a feed attractant, marine sources serve as a potential fishmeal substitute due to their rich nutritional composition. However, certain biogenic amines present in these marine-based ingredients need to be examined before their use, since these amines when present in high concentrations reduces feed intake by producing undesirable odour. Use of a combination of chemoattractants would give a better effect, rather than using them individually and compounds from plant origin have limited beneficial effects. The present review concludes that incorporation of chemoattractants would be beneficial in formulation of enriched and economical feeds with better attractability and palatability. The selection of suitable attractant and their supplementation at correct proportion is significantly more important to avoid undesirable effects in cultured shrimp. Further field-based research is needed to predict the actual effects of chemoattractants on farmed shrimp and to provide a sustainable base for the expansion of shrimp aquaculture sector, by reducing feed wastage.


Author(s):  
C. San Marchi ◽  
B. P. Somerday ◽  
X. Tang ◽  
G. H. Schiroky

Applications requiring the containment and transport of hydrogen gas at pressures greater than 70 MPa are anticipated in the evolving hydrogen economy infrastructure. Since hydrogen is known to alter the mechanical properties of materials, data are needed to guide the selection of materials for structural components. Type 316 austenitic stainless steels are often considered one of the best choices for resistance to hydrogen-assisted fracture; however, at sub-ambient temperatures some alloy compositions of type 316 stainless steel can become more susceptible to hydrogen-assisted fracture than others. In this study, we report the tensile properties of two heats of type 316 stainless steel, emphasizing the effects of temperature and high concentrations of internal hydrogen on these properties.


Entropy ◽  
2018 ◽  
Vol 20 (12) ◽  
pp. 965 ◽  
Author(s):  
Shumin Zheng ◽  
Shaoqing Wang

The elastic properties of seventy different compositions were calculated to optimize the composition of a V–Mo–Nb–Ta–W system. A new model called maximum entropy approach (MaxEnt) was adopted. The influence of each element was discussed. Molybdenum (Mo) and tungsten (W) are key elements for the maintenance of elastic properties. The V–Mo–Nb–Ta–W system has relatively high values of C44, bulk modulus (B), shear modulus (G), and Young’s modulus (E), with high concentrations of Mo + W. Element W is brittle and has high density. Thus, low-density Mo can substitute part of W. Vanadium (V) has low density and plays an important role in decreasing the brittleness of the V–Mo–Nb–Ta–W system. Niobium (Nb) and tantalum (Ta) have relatively small influence on elastic properties. Furthermore, the calculated results can be used as a general guidance for the selection of a V–Mo–Nb–Ta–W system.


1992 ◽  
Vol 8 (3) ◽  
pp. 307-314 ◽  
Author(s):  
Ricardo Ferreira Monteiro ◽  
Rogério Parentoni Martins ◽  
Kikyo Yamamoto

ABSTRACTPsittacanthus robustus (Loranthaceae) is a Neotropical mistletoe which grows mainly on species of Vochysiaceae in Brazilian ‘cerrado’ regions. Its parasitizing pattern involves: (a) the height and the aspect of the crown of the host-plants; (b) the presence of sticky substance in the fruits of the parasite; and (c) the feeding behaviour and habitat selection of the dispersal agent, the swallowtanager Tersina viridis viridis. The feeding behaviour of this bird on fruits of P. robustus facilitates seed release and seedling establishment; its foraging habit and habitat selection account for high concentrations of the parasites on their preferred host-plants.


BioResources ◽  
2020 ◽  
Vol 15 (2) ◽  
pp. 3839-3849 ◽  
Author(s):  
Yunyun Liu ◽  
Yunqi Cao ◽  
Qiang Yu ◽  
Jingliang Xu ◽  
Zhenhong Yuan

Complementary enzymes can considerably enhance the hydrolysis effectiveness of cellulase. The influence of hemicellulase supplementation on high solids saccharification of alkali-pretreated sugarcane bagasse was assessed. Hemicellulase addition of 1200 IU/g substrate with cellulase loading of 10 FPU/g substrate achieved high sugars yield with glucose and xylose conversion efficiency of 95.4% and 87.4%, respectively. To further improve the substrate conversion efficiency based on high sugars production, fed-batch hydrolysis was employed with high solids loading of 20% (w/v) to 25% (w/v). After 96 h hydrolysis with 25% solids loading at cellulase and hemicellulase loading of 20 FPU/g and 1200 IU/g substrate, respectively, the obtained highest total sugars was 242 g/L, with glucose and xylose conversion efficiencies of 98.6% and 94.9%, respectively. An increase in substrate digestibility upon supplementation of mixture enzymes with high sugars production can be realized in high solids fed-batch system with proper cellulase and hemicellulase synergism.


2020 ◽  
Vol 117 (27) ◽  
pp. 16009-16018 ◽  
Author(s):  
Ling Zou ◽  
Peter Spanogiannopoulos ◽  
Lindsey M. Pieper ◽  
Huan-Chieh Chien ◽  
Wenlong Cai ◽  
...  

Food and drug products contain diverse and abundant small-molecule additives (excipients) with unclear impacts on human physiology, drug safety, and response. Here, we evaluate their potential impact on intestinal drug absorption. By screening 136 unique compounds for inhibition of the key intestinal transporter OATP2B1 we identified and validated 24 potent OATP2B1 inhibitors, characterized by higher molecular weight and hydrophobicity compared to poor or noninhibitors. OATP2B1 inhibitors were also enriched for dyes, including 8 azo (R−N=N−R′) dyes. Pharmacokinetic studies in mice confirmed that FD&C Red No. 40, a common azo dye excipient and a potent inhibitor of OATP2B1, decreased the plasma level of the OATP2B1 substrate fexofenadine, suggesting that FD&C Red No. 40 has the potential to block drug absorption through OATP2B1 inhibition in vivo. However, the gut microbiomes of multiple unrelated healthy individuals as well as diverse human gut bacterial isolates were capable of inactivating the identified azo dye excipients, producing metabolites that no longer inhibit OATP2B1 transport. These results support a beneficial role for the microbiome in limiting the unintended effects of food and drug additives in the intestine and provide a framework for the data-driven selection of excipients. Furthermore, the ubiquity and genetic diversity of gut bacterial azoreductases coupled to experiments in conventionally raised and gnotobiotic mice suggest that variations in gut microbial community structure may be less important to consider relative to the high concentrations of azo dyes in food products, which have the potential to saturate gut bacterial enzymatic activity.


2020 ◽  
Vol 10 (1) ◽  
pp. 70-78 ◽  
Author(s):  
Zeyu Yang ◽  
Hengwei Fu ◽  
Wenjie Ye ◽  
Youyu Xie ◽  
Qinghai Liu ◽  
...  

Based on substrate-coupled cofactor regeneration system, a high 2-propanol tolerance SmADH2 together with TBCR system can synthesise structurally diverse chiral alcohols at a high substrate loading with only 1.25 equivalents of 2-propanol.


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