Biotechnology Letters最新文献

Great mass of tea tree seeds (TTS) are naturally rotted away as agricultural waste because there is no suitable technology used for extracting oil from TTS. The fermentation method is a new process and that can simultaneously recover oil, starch and saponins from TTS. In this study, a key bacterial strain named JJZ21 was isolated from fermented TTS milk (ground TTS and water) by the serial dilution method and identified as Lactiplantibacillus plantarum strain through molecular analyses. The production of cellulase, pectinase, amylase and protease of JJZ21 were visualized on agar plates containing specific enzyme substrates. Compared with the control, addition treatment with JJZ21 dramatically improved the activities of cellulase, pectinase, amylase and protease, reduced content of soluble sugar, protein and dry matter and pH in the TTS milk, shortened the fermentation time and increased the yield of TTS oil. Meanwhile, the addition of JJZ21 had no significant effect on quality of TTS oil. However, the yield of TTS oil decreased with excessive fermentation. Response surface methodology was used to evaluate the optimum conditions for fermentation process to obtain the maximum oil yield.

Plant molecular farming (PMF) has emerged as a promising strategy for producing biopharmaceuticals and high-value biomolecules in plant systems. In this review, we present a comprehensive synthesis of current methodologies while introducing novel approaches to genetic transformation, protein expression, glycan engineering, and downstream processing. We offer in-depth analyses of recent advancements such as CRISPR/Cas9-mediated pathway editing, synthetic biology frameworks for optimizing protein yield and quality, and integrated bioprocessing solutions that enhance purification efficiency. Further, detailed case studies are discussed to illustrate actionable strategies, and future research directions are proposed to bridge current gaps. By focusing on transformative techniques and critical problem-solving perspectives, this review aims to guide researchers toward more effective and scalable PMF applications.

Insect cell-baculovirus expression vector systems (IC-BEVS) are valuable tools in pharmaceutical bioprocesses for producing complex proteins like immunogenic virus-like particles. In this context, standardization is key to guarantee consistency in process yield, productivity, and product quality attributes. The study investigates the nine-month stability of the main biotechnological input used in IC-BEVS, the recombinant baculovirus vectors produced in chemically defined medium. Viral titer (pfu/mL), zeta potential (mV), and mean hydrodynamic particle size (μm), were employed to assess viral inactivation under eight combinations of generation and storage conditions. First-order, Weibull, and biphasic models were applied to describe viral decay. The critical parameters (factors) analyzed were the size of the heterologous gene inserted (719 and 1621 bp), storage temperature (- 80 and 1.5 °C), and infection time used for baculovirus batch generation (48 or 72 h post-infection). They were mainly explored according to a two-level factorial design (2³). The primary quality attribute evaluated in this study was the one-log₁₀ decay time of the viral titer (td), which exhibited an overall mean value of 80 days across eight batches. The biphasic model best fits the dispersion of the viral titer data collected over the assessed time in all considered combinations of factors and was employed to find significant factors over td values. Gene size was the only factor with a statistically significant effect on viral titer decay; additionally, the study indicates the occurrence of particle aggregation over the course of the analysis.

D-Glucosaminic acid is a valuable amino acid useful in food and medical applications. It is a highly sought after enantiopure molecule important for the synthesis of drugs and glycopeptides. Current enzymatic synthesis pathways to D-glucosaminic acid carry disadvantages such as low product yield and long reaction times. Herein, the Auxiliary Activity 7 chito-oligosaccharide oxidase from Lentinus brumalis, LbChi7A, was shown as a potent biocatalyst capable of efficiently converting D-glucosamine (GlcN) and N-acetyl-D-glucosamine (GlcNAc) to their respective C₁-acids. The substrate specificity of LbChi7A towards GlcN and GlcNAc enabled the conversion of at least 90% GlcN to D-glucosaminic acid and 100% GlcNAc to N-acetyl-D-glucosaminic acid within 60 min. LbChi7A inhibition by the hydrogen peroxide co-product was not detected, even at 860 mM. This single enzymatic conversion offers a clean and efficient process to produce industrially relevant glucosaminic acids, including D-glucosaminic acid and N-acetyl-D-glucosaminic acid.

Xylanases have attracted considerable attention due to their strong potential for industrial use. In this study, a xylanase-producing strain isolated from soil was identified as Trichoderma semiorbis Tsejk8, and the conditions for xylanase production were optimized. Additionally, two xylanase-related genes were cloned, and their functions were analyzed. The optimal conditions for xylanase production included maltose as the carbon source, peptone as the nitrogen source, an optimal pH of 6.0, and an incubation time of 120 h, yielding an enzyme activity of 40.7 U/mL. Following the purification of xylanase via ammonium sulfate precipitation and ion exchange chromatography, four distinct protein bands were observed. Mass spectrometry analysis of these bands identified 14 associated proteins. Bioinformatics analysis revealed that two of these proteins belong to GH3 (Glycoside Hydrolase family 3) β-xylosidase. In summary, the newly isolated strain Tsejk8 exhibits xylanase activity, offering an effective and eco-friendly means of converting biomass into raw materials for industrial applications.

The aldehyde dehydrogenase (ALDH) superfamily plays a critical role in acetaldehyde detoxification. The mitochondrial ALDH2 isoenzyme serves as the core enzyme in alcohol metabolism because of its high affinity for acetaldehyde. Loss-of-function mutation (such as ALDH2*2) of the gene is highly prevalent in East Asian populations. These mutations cause acetaldehyde accumulation and significantly increase the risk of alcohol-related diseases. Therefore, it is necessary to develop efficient recombinant ALDH2 supplementation therapies. However, its heterologous expression faces three major bottlenecks: (1) catalytic turnover leading to NAD⁺ depletion; (2) difficulty in functional tetramer assembly; and (3) cytotoxicity of the substrate acetaldehyde, which inhibits cell growth. The review covers rational host selection and multi-level synergistic optimization of transcription, translation, folding, and metabolism. Examples include promoter optimization, codon optimization, terminator optimization, and chaperone co-expression. Ultimately, we explore essential stabilization formulations and innovative delivery strategies, such as nano-encapsulation and engineered probiotics, needed to realize the therapeutic potential of ALDH2. The goal is to promote the industrial production and clinical translation of recombinant ALDH2.

Chemical surface modification of enzymes represents a powerful strategy to improve immobilization and catalytic performance. In this study, glucose oxidase (GOx) from Aspergillus niger was chemically modified via periodate oxidation and reductive amination to incorporate L-aspartate (D-GOx) and L-histidine (H-GOx) residues. These modifications enhanced electrostatic and coordinative interactions with Zn²⁺ and 2-methylimidazole during biomimetic mineralization, leading to efficient encapsulation in ZIF-8 frameworks. The resulting D-GOx@ZIF-8 and H-GOx@ZIF-8 biocomposites showed significantly improved activity and stability compared to native GOx. H-GOx@ZIF-8 achieved a specific activity of 4551 IU/g, representing a sixfold increase over previously reported systems. Both modified biocomposites also demonstrated greater resistance to detergent washing and retained higher activity after exposure to 65 °C, indicating stronger incorporation into the ZIF-8 matrix. These results highlight the dual advantage of introducing negatively charged and imidazole-functional groups for promoting biomineralization and improving biocatalyst robustness. This strategy provides a mild, scalable route for preparing enzyme@MOF composites with enhanced operational stability, offering direct potential for industrial bioprocesses, continuous-flow catalysis, and biosensing applications.

Purpose: This narrative review aims to summarize current evidence on antisense oligonucleotide (ASO)-based strategies targeting oncogenic long non-coding RNAs (lncRNAs) and to evaluate their therapeutic potential in cancer initiation, progression, metastasis, and treatment resistance.

Methods: A narrative review of preclinical studies was conducted, focusing on ASO-mediated silencing of cancer-associated lncRNAs, including MALAT1, PVT1, NEAT1, and other emerging lncRNAs, across multiple malignancies such as lung, breast, ovarian, gastrointestinal, and head-and-neck cancers. Advances in ASO chemical design and delivery strategies-including lipid and polymeric nanoparticles, ligand-directed conjugates, peptide-based carriers, and gymnotic uptake-were also examined.

Results: Preclinical evidence consistently demonstrates that lncRNA-targeted ASOs effectively suppress tumor growth, invasion, metastasis, and chemoresistance while promoting apoptosis. Improvements in ASO chemical modifications and delivery platforms have significantly enhanced ASO stability, biodistribution, and intracellular uptake, thereby improving therapeutic efficacy across diverse cancer models.

Conclusion: Targeting oncogenic lncRNAs using ASO-based approaches represents a promising and rapidly evolving strategy in precision oncology. Although substantial progress has been achieved in ASO design and delivery, further optimization and well-structured clinical trials are required to facilitate the translation of these therapies into routine cancer treatment.

Vaccination remains one of the effective strategies to control infectious diseases, thereby preventing millions of deaths each year globally. Veterinary vaccines protects animal and public health, enabling efficient production of livestock, reduce antibiotic use and also prevent zoonotic spill over events. Traditional vaccines were developed using live-attenuated or inactivated pathogens. However, the advancements in the genetic engineering and recombinant protein production technologies have made possible the vaccine design and production of recombinant vaccine antigens or immunotherapeutic molecules for both human and veterinary use. Most of the commercially available recombinant therapeutic proteins are produced in mammalian system which limits their feasibility, particularly for veterinary applications due to their high production costs. Therefore, the development of low cost products using alternative production platforms is highly essential. The utilization of plant expression system for the recombinant vaccine production has increased in the recent years due to its advantages such as low production costs, scalability, safety, and the ability to express complex proteins. This review provides a comprehensive overview of the vaccine antigens produced in plant system with a particular focus on vaccines targeting infectious diseases in ruminants and swine.

Developing stable and easy-to-operate biocatalysts is crucial for their use as industrial catalysts. Here immobilized whole-cell catalysts were used for β-alanine production by immobilizing recombinant Escherichia coli cells (containing hydroaminase) with diatomite. E. coli BL21 (DE3)/pET-30a ( +)-HAMase was genetically engineered for the efficient synthesis of β-alanine from acrylic acid and aqueous ammonia. Using glutaraldehyde as a cross-linking agent, polyethyleneimine (PEI) as a flocculant, and diatomite as the immobilization carrier, optimal immobilization was achieved with 8% (w/v) PEI solution, 5% (w/v) glutaraldehyde, and 100 mg wet cell/mL cell suspension, along with a PEI flocculation time of 2.5 h and glutaraldehyde cross-linking time of 1.5 h. The enzyme activity recovery rate reached 70.72%. Remarkably, the immobilized whole-cell catalysts exhibited excellent stability, retaining over 90% of initial enzyme activity after 12 h incubation at 45 °C and maintaining over 72% enzyme activity after storage for 60 days at 4 °C. Additionally, the immobilized cells demonstrated enhanced reusability, maintaining consistent β-alanine yield even after ten consecutive reaction batches with an average yield of approximately 80%.