Preparative Biochemistry & Biotechnology最新文献

In this study, biocatalytic transesterification reaction using Novozyme 435^® (N435) lipase was employed to enhance the hydrophobicity of esculin, aiming to improve its solubility for commercial applications and enhance its bioactivity and oral viability. The acylation reaction of esculin with vinyl acetate was conducted at 60 °C and 200 rpm for 24 h. After chromatographic and spectroscopic analysis, two products were identified: the first one was monoacylated at the 6'-OH position of the glucosyl moiety of esculin (T_R: 10.3 min and m/z 382.93 [M + H]⁺), and the second one was diacylated at the 6'-OH and 3'-OH positions (T_R: 13.0 min and m/z 424.93 [M + H]⁺). The latter was the major product, with a conversion rate of 53.550 ± 0.368%, while the monoacetylated one showed 8.715 ± 0.064%. Both products were isolated by high-speed counter-current chromatography (HSCCC) using a two-phase system HEMWat 1:9:1:9 and characterized by NMR. In this way, these results improve the practical application of esculin, through the obtention of esculin mono and diacetates by fast and efficient biocatalysis reaction.

Bletilla striata polysaccharide (BSP) is one of the main active ingredients of the traditional Chinese medicine Bletilla striata (Thunb) Richb.f and the extraction method of BSP has a significant impact on its properties. This study investigated the effects of four extraction methods, namely hot water extraction, ultrasonic extraction, enzyme extraction, and microwave extraction, on the structure and antioxidant properties of BSP. Characterization results from FTIR and NMR showed that all four BSP consisted mainly of glucose and mannose, forming α-glycosidic and β-glycosidic linkages to form glucomannan. Hot water extraction had the lowest extraction rate of BSP at 21.78% ± 0.73%. The polysaccharide BSP-H obtained from hot water extraction had the smallest absolute Zeta potential and Grain size, but the largest molecular weight at 204 kDa. It exhibited the best thermal stability and superior antioxidant activity compared to polysaccharides extracted using the other three methods, as evaluated by three different antioxidant assays. Although the antioxidant activity of BSP-V was slightly weaker, it showed a significant improvement compared to the remaining two polysaccharides. These results suggest that hot water extraction is the most suitable method for large-scale application of BSP, preserving its activity effectively, thus facilitating practical production and product development.

The present study reports the green synthesis of cellulose nanocrystals from the shells of Sterculia foetida (SFS) cellulose. Three different methods, alkali, acid and organic acid, were screened for the maximum cellulose extraction. A maximum cellulose yield, 30.6 ± 0.84 w/w, was obtained using 90% formic acid at 110 °C in 120 min. The extracted cellulose was characterized and identified by instrumental analyses. SEM analysis showed skeletal rod-like microfibril structures and similar intra-fibrillar widths. CP/MAS ¹³C NMR and FTIR spectrum revealed the purity of cellulose and the absence of other components like hemicellulose and lignin. XRD study revealed a cellulose crystallinity index of 88.07%. BET analysis showed a good surface area (3.3213 m²/g) and a micro-pore area of 1.871 m²/g. The cellulose nanocrystals were synthesized from the extracted cellulose using deep eutectic solvents (DES), choline chloride and lactic acid (1:2 ratio). The cellulose nanocrystals (CNC) synthesized from DES-based exhibited zeta potential and particle size of -16.7 mV and 576.3 d.nm. DES-synthesized cellulose nanocrystals were spherical-like shapes, as observed from TEM images. The present results exposed that formic acid is an effective and green catalyst for the extraction of cellulose and DES for the sustainable synthesis of CNC.

Calendula officinalis is a widespread medicinal plant with a sufficiently well-studied chemical composition. Secondary metabolites synthesized by C.officinalis plants have pharmacological value for treating numerous diseases, and various types of aseptic in vitro cultures can be used as a source of these compounds. From this perspective, hairy roots attract considerable attention for the production of bioactive chemicals, including flavonoids with antioxidant activity. This paper shows the possibility of C.officinalis hairy roots obtaining with 100% frequency by Agrobacterium rhizogenes genetic transformation. Hairy root lines differed in growth rate and flavonoid content. In particular, flavonoids were accumulated in the amount of up to 6.68 ± 0.28 mg/g of wet weight. Methyl jasmonate in the concentration of 10 µM inhibited root growth to a small extent but stimulated the synthesis of flavonoids. The antioxidant activity and the reducing power increased in the roots grown in the medium with methyl jasmonate. The strong correlation of antioxidant activity and reducing power with flavonoid content was detected. The influence of extraction conditions on the content of flavonoids in the extracts and their bioactivity was determined. The potent reducing activity of extracts from hairy roots allowed the production of silver nanoparticles, which was confirmed by transmission electron microscopy.

In recent years, driven by increasing consumer demand for natural and healthy convenient foods, the food industry has been shifting from synthetic to natural products. This shift is also reflected in the growing popularity of non-conventional extraction methods for pigments, which are favored for sustainability and environment-friendliness compared to conventional processes. This review aims to investigate the extraction of carotenoids from a variety of natural sources, including marine sources like fungus, microalgae, and crustaceans, as well as widely studied plants like tomatoes and carrots. Additionally, it delves into the recovery of valuable carotenoids from waste products like pomace and peels, highlighting the nutritional and environmental benefits. The review also emphasizes the role of green solvents such limonene, vegetable oils, ionic liquids, supercritical fluids, and natural deep eutectic solvents in effective and ecologically friendly carotenoid extraction. These technologies support the ideas of a circular and sustainable economy in addition to having a smaller negative impact on the environment. Overall, the present study highlights the crucial importance of green extraction technologies in achieving the dual goals of sustainability and public safety.

Botrytis cinerea, an airborne plant pathogen, holds the potential to synthesize sesquiterpenes, which have been used for the industrial production of abscisic acid. Previously, through our genetic technology, we obtained strain ZX2, whose main product 1´,4´-trans-ABA-diol is physiologically active in plants. In this study, 50 L of fed fermentation was carried out with ZX2 strain to study the stability of expression of TUA, TUB, ATC, EF-1, GAPDH, UCE and GTP genes. Four kinds of software (GeNorm, NormFinder, BestKeeper and Delta Ct) were used to analyze the expression stability of candidate genes, and finally the best reference gene was screened by RefFinder. Based on the results, the ACT was the most stable gene. It was used to normalize the expression levels of two genes related to 1´,4´-trans-ABA-diol production (hmgr and bcaba3) when fed-batch fermentation. Guide the selection of appropriate internal reference genes during the fermentation process to accurately quantify the relative transcription levels of target genes in B.cinerea ZX2.

Investigating the biotechnological potential of wild microorganisms is paramount for optimizing bioprocesses. Given this premise, we looked for yeasts in Brazilian native stingless bees, considering the recognized potential of pollinating insect-associated microorganisms for the production of volatile organic compounds (VOCs). Two yeast strains of the species Meyerozyma caribbica were isolated from bees Scaptotrigona postica and evaluated for their fermentative capacity. Both yeasts were capable of fermenting sucrose (the main sugar used in the Brazilian ethanol industry) with over 90% efficiency and yields of up to 0.504 g/g. Through an experimental design analysis (CCD), it was verified that the ethanol productivity of these yeasts can also benefit from high concentrations of sucrose and low pH values, desirable traits for microorganisms in this biofuel production. At the same time, CCD analyses also showed the great capacity of these M. caribbica strains to produce another alcohol of broad biotechnological interest, 2-phenylethanol. Interestingly, the statistical analyses demonstrated that greater production of this compound can occur at high sugar concentrations and low availability of nitrogen sources, which can be easily achieved using residual low-cost feedstocks. Thus, our results suggest that these M. caribbica strains may be efficiently used in both ethanol and 2-phenylethanol production.

This study investigated the ultrasound-assisted extraction (UAE) techniques used to enhance the polyphenols and antioxidants of mango peel extract (MPE). Additionally, it explored the bacteriostatic activity of MPE against various microorganisms. The UAE method was optimized using response surface methodology (RSM) at different times, temperatures, and ratios, with optimal conditions found to be 35 minutes, 45 °C, and a 1:35 ratio. The optimized yield results for total polyphenol content (TPC) were 17.33 ± 1.57 mg GAE/g, total flavonoid content (TFC) was 12.14 ± 0.29 mg QE/g, and radical scavenging activity (RSA) was 72.11 ± 2.19%. These response models were extremely significant with p-values less than 0.05. MPE showed selective effectiveness against Bacillus cereus, Geobacillus stearothermophilus, and Escherichia coli (E. coli). The results highlight the potential of mango peel as a sustainable source of bioactive compounds, contributing to waste reduction in the food industry and the development of natural antimicrobial agents. This study contributes to further research on the application of MPE in processed foods.

Traditional Chinese medicine (TCM) is often composed of a variety of natural medicines. Its composition is complex, and many of its components can not be analyzed and identified. The first step in the rational application of TCM is to successfully separate the effective components which is also a great inspiration for the development of new drugs. Among the many separation technologies of TCM, the traditional heating concentration separation technology has high energy consumption and low efficiency. As a new separation technology, membrane separation technology has the characteristics of simple operation, high efficiency, environment-friendly and so on. The separation effect of high molecular weight difference solution is better. The applications of several main membrane separation technologies such as microfiltration, nanofiltration, ultrafiltration and reverse osmosis are reviewed, the methods of restoring membrane flux after membrane fouling are discussed, and their large-scale industrial applications in the future are prospected and summarized.

In the [Bmim]Cl reaction medium, five different acidic ionic liquids were used as catalysts to study the effects of reaction time, reaction temperature, system water content, catalyst dosage, microwave power, and other factors on cellulose hydrolysis under microwave irradiation. The results showed that in the [Bmim]Cl reaction system, using N-methylpyrrolidone methylsulfonic acid salt as a catalyst, controlling the microwave reaction time at 10 min, reaction temperature at 130 °C, catalyst dosage at 1 g/g (cellulose), water addition at 0.756 μL/g ([Bmim]Cl), and microwave power at 480 W, resulted in the best cellulose hydrolysis effect with a glucose yield of 74.49%. Compared to conventional heating, the glucose yield increased by 24% and the hydrolysis time was reduced by 77%. Microwave irradiation significantly enhances the cellulose hydrolysis process in an ionic liquid medium.