Microbacterium salsuginis sp. nov., a halotolerant actinobacterium with antimicrobial activity.

A novel Gram-staining-positive actinobacterium with antimicrobial activity, designated CFH 90308T, was isolated from the sediment of a salt lake in Yuncheng, Shanxi, south-western China. The isolate exhibited the highest 16S rRNA gene sequence similarities to Microbacterium yannicii G72T, Microbacterium hominis NBRC 15708T and Microbacterium xylanilyticum S3-ET (98.5, 98.4 and 98.2 %, respectively), and formed a separate clade with M. xylanilyticum S3-ET in phylogenetic trees. The strain grew at 15-40 ºC, pH 6.0-8.0 and could tolerate NaCl up to a concentration of 15 % (w/v). The whole genome of strain CFH 90308T consisted of 4.33 Mbp and the DNA G+C content was 69.6 mol%. The acyl type of the peptidoglycan was glycolyl and the whole-cell sugars were galactose and mannose. The cell-wall peptidoglycan mainly contained alanine, glycine and lysine. The menaquinones of strain CFH 90308T were MK-12, MK-13 and MK-11. Strain CFH 90308T contained anteiso-C15:0, anteiso-C17:0, iso-C16:0 and iso-C15:0 as the predominant fatty acids. The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between CFH 90308T and the other species of the genus Microbacterium were found to be low (ANIb <81.3 %, dDDH <25.6 %). The secondary metabolite produced by strain CFH 90308T showed antibacterial activities against Bacillus subtilis, Pseudomonas syringae, Aeromonas hydrophila and methicillin-resistant Staphylococcus aureus. Based on genotypic, phenotypic and chemotaxonomic results, the isolate is considered to represent a novel species of the genus Microbacterium, for which the name Microbacterium salsuginis sp. nov. is proposed. The type strain is CFH 90308T (=DSM 105964T=KCTC 49052T).

Improving the Catalytic Properties of Xylanase from Alteromones Macleadii H35 Through Sequence Analysis.

Endo-1,4-ß-xylanase is a key xylanolytic enzyme, and our study aimed to enhance the catalytic properties of Alteromones Macleadii xylanase (Xyn ZT-2) through sequence-guided design approach. Analysis of the amino acid sequence revealed highly conserved residues near the active site, with few differences. Introducing various mutations allowed us to modify the enzyme's catalytic performance. Particularly, the A152G mutation led to a 9.8-fold increase in activity and a 23.2-fold increase in catalytic efficiency. Moreover, A152G exhibited an optimal temperature of 65 °C, 20 °C higher than that of Xyn ZT-2, while the T287S mutant showed a 4.9-fold increase in half-life. These results underscore the role of amino acid evolution in shaping xylanase catalysis. Through targeted sequence analysis and a focused mutation library, we effectively improved catalytic performance, providing a straightforward approach for enhancing enzyme efficiency.

One-pot biosynthesis of gastrodin using UDP-glycosyltransferase itUGT2 with an in situ UDP-glucose recycling system.

Gastrodin, the major effective ingredient in Tianma (Gastrodia elata), is a p-hydroxybenzoic acid derivative with various activities. Gastrodin has been widely investigated for food and medical applications. The last biosynthetic step for gastrodin is UDP-glycosyltransferase (UGT)-mediated glycosylation with UDP-glucose (UDPG) as glycosyl donor. In this study, we performed a one-pot reaction both in vitro and in vivo to synthesize gastrodin from p-hydroxybenzyl alcohol (pHBA) by coupling UDP-glucosyltransferase from Indigofera tinctoria (itUGT2) to sucrose synthase from Glycine max (GmSuSy) for regeneration of UDPG. The in vitro results showed that itUGT2 transferred a glucosyl group to pHBA to generate gastrodin. After 37 UDPG regeneration cycles with 2.5% (molar ratio) UDP, the pHBA conversion reached 93% at 8 h. Furthermore, a recombinant strain with itUGT2 and GmSuSy genes was constructed. Through optimizing the incubation conditions, a 95% pHBA conversion rate (220 mg/L gastrodin titer) was achieved in vivo without addition of UDPG, which was 2.6-fold higher than that without GmSuSy. This in situ system for gastrodin biosynthesis provides a highly efficient strategy for both in vitro gastrodin synthesis and in vivo biosynthesis of gastrodin in E. coli with UDPG regeneration.

Research progress on the mechanism of chronic neuropathic pain.

Chronic neuropathic pain (CNP) refers to pain that lasts for more than three months due to a disease or an injury to the somatosensory nervous system. The incidence of CNP has been increasing in the world, causing it to become a global concern and patients often experience spontaneous pain, hyperalgesia, abnormal pain or even abnormal sensation as some of its main symptoms. In addition to serious pain and poor physical health, CNP also negatively affects patients' mental health, thus impacting the overall quality of their lives. The pathogenesis of CNP is not clear, but some studies have proved that central sensitization, peripheral sensitization, neuroinflammation, dysfunction in descending nociceptive modulatory systems, oxidative stress reaction, activation of glial cells and psychological factors play an important role in the occurrence and development of CNP. In this context, this article summarizes the current research progress on the mechanism of CNP to provide a basis for further research in preventing and treating the disease.

Cardiac magnetic resonance radiomics for disease classification.

OBJECTIVES: This study investigated the discriminability of quantitative radiomics features extracted from cardiac magnetic resonance (CMR) images for hypertrophic cardiomyopathy (HCM), dilated cardiomyopathy (DCM), and healthy (NOR) patients. METHODS: The data of two hundred and eighty-three patients with HCM (n = 48) or DCM (n = 52) and NOR (n = 123) were extracted from two publicly available datasets. Ten feature selection methods were first performed on twenty-one different sets of radiomics features extracted from the left ventricle, right ventricle, and myocardium segmented from CMR images in the end-diastolic frame, end-systolic frame, and a combination of both; then, nine classical machine learning methods were trained with the selected radiomics features to distinguish HCM, DCM, and NOR. Ninety classification models were constructed based on combinations of the ten feature selection methods and nine classifiers. The classification models were evaluated, and the optimal model was selected. The diagnostic performance of the selected model was also compared to that of state-of-the-art methods. RESULTS: The random forest minimum redundancy maximum relevance model with features based on LeastAxisLength, Maximum2DDiameterSlice, Median, MinorAxisLength, Sphericity, VoxelVolume, Kurtosis, Flatness, and Skewness was the highest performing model, achieving 91.2% classification accuracy. The cross-validated areas under the curve on the test dataset were 0.938, 0.966, and 0.936 for NOR, DCM, and HCM, respectively. Furthermore, compared with those of the state-of-the-art methods, the sensitivity and accuracy of this model were greatly improved. CONCLUSIONS: A predictive model was proposed based on CMR radiomics features for classifying HCM, DCM, and NOR patients. The model had good discriminability. KEY POINTS: • The first-order features and the features extracted from the LOG-filtered images have potential in distinguishing HCM patients from DCM patients. • The features extracted from the RV play little role in distinguishing DCM from HCM. • The VoxelVolume of the myocardium in the ED frame is important in the recognition of DCM.

Cloning, Expression, and Characterization of a GHF 11 Xylanase from Alteromonas macleodii HY35 in Escherichia coli.

A xylanase gene xynZT-1 from Alteromonas macleodii HY35 was cloned and expressed in Escherichia coli (E. coli). The sequencing results showed that the ORF of xynZT-1 was 831 bp. The xylanase DNA sequence encoded a 29 amino acids (aa) signal peptide and a 247-aa mature peptide. The XynZT-1 has been a calculated molecular weight (MW) of 27.93 kDa, isoelectric point (pI) of 5.11 and the formula C1266H1829N327O384S5. The amino acid sequence of the xynZT-1 had a high similarity with that of glycosyl hydrolase family 11 (GHF11) reported from other microorganisms. The DNA sequence encoding mature peptide was subcloned into pET-28a(+) expression vector. The resulted plasmid pET-28a-xynZT-1 was transformed into E. coli BL21(DE3), and the recombinant strain BL21(DE3)/xynZT-1 was obtained. The optimum temperature and pH of the recombinant XynZT-1 were 45 ℃ and 5.0, respectively.

Characterization of polyphosphate kinases for the synthesis of GSH with ATP regeneration from AMP.

Polyphosphate kinase (PPK) is important for industrial processes involving ATP regeneration. While a variety of methods have been reported for regenerating ATP from ADP, few have explored enzyme catalyzed ATP regeneration from cheaper and stable AMP. In this work, PPKs from different sources were expressed and their catalytic activity were tested at different reaction temperatures, reaction pH and with different polyphosphate (polyPn) types. The ATP regeneration system for glutathione (GSH) synthesis was established using a single PPK capable of phosphorylating AMP to synthesize ATP from AMP and short chain polyPn. GSH yield was obtained using adenosine mono-, di- and triphosphates, which confirmed the flexibility of our constructed ATP regeneration system coupled with GSH synthesis via bifunctional GSH synthase. Finally, optimization of the GSH synthesis yielded conversion value above 80 %. Overall, these results illustrate that PPK is suitable for a broader range of substrates than previously expected, and has great untapped potential for applications involving ATP regeneration.

Brachybacterium subflavum sp. nov., a novel actinobacterium isolated from the foregut of grass carp.

A novel actinobacterium, designated CFH 10395T, was isolated from the foregut of grass carp (Ctenopharyngodon idella), which had been fed with ginseng extract supplement. The taxonomic position was investigated by a polyphasic approach. Cells of CFH 10395T were Gram-staining-positive, aerobic, ovoid-shaped, non-spore-forming and non-motile. On the basis of the results of 16S rRNA gene sequence analysis, CFH 10395T was most closely related to Brachybacterium endophyticum KCTC 49087T, Brachybacterium squillarum JCM 16464T and Brachybacterium paraconglomeratum JCM 17781T (97.85%, 97.51 and 97.29% similarity, respectively). CFH 10395T grew at 4-37 °C, pH 5.0-9.0 and in the presence of up to 10.0 % NaCl (w/v). The dominant menaquinone was MK-7. The whole-cell sugars were rhamnose, glucose, mannose and galactose. meso-diaminopimelic acid was the diagnostic diamino acid in the cell-wall peptidoglycan. The major fatty acids were anteiso-C15 : 0, anteiso-C17 : 0 and iso-C16 : 0. The genome size was 3.99 Mbp with a DNA G+C content of 71.9 mol%. On the basis of the results of phylogenetic analysis, physiological properties, chemotaxonomic characteristics, low average nucleotide identity (ANI) and digital DDH (dDDH) results [ANI calculated using MUMmer (ANIm) <87 %, ANI calculated using blast (ANIb) <83 % and dDDH <23 %], it is concluded that CFH 10395T represents a novel species of the genus Brachybacterium, for which the name Brachybacterium subflavum sp. nov., is proposed. The type strain is CFH 10395T (=CGMCC 1.13804T=KCTC 49235T).

Immobilization of laccase by 3D bioprinting and its application in the biodegradation of phenolic compounds.

Due to the increasing quantities of phenolic compounds present in wastewater, the use of enzymatic degradation with the laccase has attracted much attention as a green option for their removal. In this work, we developed a novel immobilization technology using 3D bioprinting for laccase immobilization. The hydrogel mechanism properties were optimized by experimenting with different component ratios of sodium alginate (SA), acrylamide (AM), and hydroxyapatite (HA). The improved mechanism properties were validated by morphology pictures and rheology characteristics. The optimal AM:HA:SA ratio was determined to be 4:1.2:1. We then employed an extrusion-based bioprinting technique to prepare the immobilized laccase. The substrate conversion was increased with the addition of HA, which improved the permeability of the matrix, and proved to be suitable for immobilization. The resulting immobilized laccase was used for the biodegradation of p-chlorophenol. The effects of the initial substrate concentration, pH, and temperature were evaluated. The immobilized laccase exhibited good storage stability and reusability, retaining over 80% of its initial activity after 72 h of storage, and was able to be reused for seven batches. These results highlight that the immobilized laccase prepared by 3D bioprinting has great potential for use in the biodegradation of phenolic compounds.

Aliiroseovarius marinus sp. nov., isolated from seawater.

A novel Gram-stain-negative, rod-shaped, non-spore-forming, non-flagellated, strictly aerobic strain, designated A6024T, was isolated from coastal seawater near Rizhao, PR China (119.61° E 35.47° N). The organism grew optimally at 28 °C, in pH 6.0-7.0 and in the presence of 3.0 % (w/v) NaCl. The strain required seawater or artificial seawater for growth and NaCl alone did not support growth. Strain A6024T contained ubiquinone 10 as the sole respiratory quinone and C18 : 1 ω7c (75.2 %) as the most abundant fatty acid. The predominant polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylcholine, one unidentified aminolipid and two unidentified lipids. The DNA G+C content of strain A6024T was 59.9 mol%. The results of phylogenetic analysis based on 16S rRNA gene sequences showed that the novel strain was related most closely to Aliiroseovarius halocynthiae MA1-10T, Aliiroseovarius pelagivivens GYSW-22T and Aliiroseovarius crassostreae CV919-312T with 98.3, 97.6 and 97.4 % sequence similarities, respectively. The calculated average nucleotide identity values and DNA-DNA hybridization values between strain A6024T and the phylogenetically related Aliiroseovarius species were in the range 76.0-85.6 % and 19.6-29.4 %, respectively. On the basis of the results of polyphasic analyses, strain A6024T represents a novel species of the genus Aliiroseovarius, for which the name Aliiroseovarius marinus sp. nov. is proposed. The type strain is A6024T (=KCTC 72114T=MCCC 1K03595T).

Oncolytic viro-chemotherapy exhibits antitumor effect in laryngeal squamous cell carcinoma cells and mouse xenografts.

Background: Oncolytic virus can specifically replicate in and then lyse tumor cells, but seldom in normal cells. Further studies have shown the significant therapeutic effect of oncolytic virotherapy combining with other strategies, such as chemo-, radio-, and immunotherapy et al. In this study, we investigated the combinational effect of oncolytic virus ZD55-TRAIL and chemotherapy drug doxorubicin (DOX) on human laryngeal squamous cell carcinoma (LSCC). Methods: The effect of ZD55-TRAIL combined with DOX on cell growth was assessed in LSCC Hep2 cells and normal cells by MTT assay. Hochest 33342 staining was performed to observe cell morphological changes. Western blot was used to detect the expression of apoptotic activation proteins. The in vivo antitumor efficacy of combination treatment was estimated in laryngeal cancer xenograft models. Results: The combination of ZD55-TRAIL and DOX exhibited enhanced inhibitory effects on laryngocarcinoma cell growth, and had few side effects to normal cells in vitro. Chemotherapy drug increased the inducement of tumor cell apoptosis mediated by oncolytic virus. In vivo experiment confirmed that the combination treatment significantly inhibited Hep2 laryngocarcinoma xenografts growth in mice. Conclusion: The oncolytic viro-chemotherapy is a potent therapeutic approach for in vitro cytotoxicity evaluation of Hep2 cells and xenograft growth in vivo.

Improvement of lipase activity by synergistic immobilization on polyurethane and its application for large-scale synthesizing vitamin A palmitate.

We have developed an improved and effective method to immobilize lipase on hydrophobic polyurethane foam (PUF) with different modifications. PUF was treated with hydrochloric acid to increase the active sites and then the active carboxyl groups and amino groups were exposed. Enzyme activity of lipase immobilized on PUF-HCL (8000 U/g) was 50% higher than that of lipase immobilized on PUF (5300 U/g). There is an increase in the activity of the immobilized lipase on AA/PEI-modified support (115,000 U/g), a 2.17-fold increase compared to lipase immobilized on the native support was observed. The activity of immobilized lipases was dependent on the PEI molecular weight, with best results from enzyme immobilized on PUF-HCL-AA/PEI (MW 70,000 Da, 12,800 U/g)), which was 2.41 times higher compared to that of the same enzyme immobilized on PUF. These results suggest that the activity of immobilized lipase is influenced by the support surface properties, and a moderate support surface micro-environment is crucial for improving enzyme activity. Finally, the immobilized lipase was used for the production of vitamin A palmitate. The immobilized lipase can be reused for up to 18 times with a conversion rate above 90% for 12 h in a 3 L bioreactor. Research highlights An efficient immobilization protocol on polyurethane foam was developed Polyethyleneimine and acetic acid were used to regulate the micro-environment concurrently The activity of lipase immobilized on PUF-HCL-AA/PEI was improved by 2.41 times Immobilized lipase exhibited excellent operational stability for vitamin A palmitate synthesis.

MicroRNA-625 inhibits cell invasion and epithelial-mesenchymal transition by targeting SOX4 in laryngeal squamous cell carcinoma.

INTRODUCTION: Laryngeal squamous cell carcinoma (LSCC) is a highly aggressive malignant cancer, but the molecular mechanisms underlying its development and progression remain largely elusive. The purpose of the present study is to investigate the expression profile and functional role of microRNA-625 (miR-625) in LSCC. MATERIALS AND METHODS: LSCC tissues and adjacent normal tissues were collected from 86 LSCC patients. The expression levels of miR-625 and SOX4 mRNA in tissues and cells were detected by RT-qPCR analysis. The expression levels of SOX4 and EMT-related proteins were detected by western blot analysis. In vitro cell proliferation, migration, and invasion were detected by MTT assay, colony formation assay, wound healing assay, and transwell invasion assay, respectively. Dual-luciferase reporter assay was performed to verify the binding relationship between miR-625 and the 3'-UTR of SOX4. RESULTS: The results demonstrated that miR-625 is significantly down-regulated in clinical LSCC tissues, and its low expression may be closely associated with unfavorable clinicopathological characteristics of LSCC patients. Overexpression of miR-625 significantly suppressed the proliferation, migration, invasion, and EMT of LSCC cells. Furthermore, SOX4 was validated as a direct target of miR-625 in LSCC cells, and rescue experiments suggested that restoration of SOX4 blocked the tumor suppressive role of miR-625 in LSCC cells. CONCLUSIONS: Taken together, these findings highlighted a critical role of miR-625 in the pathogenesis of LSCC, and restoration of miR-625 could be considered as a potential therapeutic strategy against this fatal disease.

Lipase immobilization on high water adsorbing capacity bagasse: applications in bio-based plasticizer synthesis.

This study investigates the structure and water adsorbing capacity of bagasse and of sodium hydroxide pretreated bagasse. The structures of bagasse and bagasse-NaOH were compared by SEM and XRD. Candida antarctica lipase B was then immobilized on bagasse, bagasse-NaOH and DPA@bagasse-NaOH. The expressed activity and immobilization yield of lipase immobilized on bagasse-NaOH (1.0%) was 36% and 45% higher than that on bagasse. When dopamine (DPA) was used as cationic polymer monomer via self-polymerization for mediating immobilization, the protein loading amounts and activity of lipase immobilized on DPA@bagasse-NaOH were higher than that of bagasse-NaOH. When the DPA concentration was 100 mg/ml, the immobilized lipase expressed activity reached its highest value (800 U/g), where the immobilization yield achieved 96.8%, which was 3.93-fold of lipase immobilized on native bagasse (24.6%). Then the immobilized lipases were used to synthesize a bio-based plasticizer. Lipase immobilized on DPA@bagasse-NaOH exhibited a significantly improved operational stability. Even after 12 batches, a high ester yield (84.2%) was maintained. Additionally, poly (vinyl chloride) PVC blends plasticized with methyl oleate as a secondary plasticizer were investigated. It was discovered that methyl oleate can be used as an effective bio-based plasticizer for PVC. These results indicate that bagasse with high water adsorbing capacity and self-polymerized DPA layer could create a favorable microenvironment for bio-based plasticizer synthesis in esterification reactions.

Environmentally-friendly strategy for separation of 1,3-propanediol using biocatalytic conversion.

Glycerol waste from the biodiesel production can be used as a carbon source in the production of 1,3-propanediol (1,3-PD) through microbial fermentation. However, downstream processing is a major bottleneck that restricts its biological production. Here, we investigated an environmentally-friendly method to enzymatically separate 1,3-PD. The transformation of 1,3-PD to an ester was achieved by exploiting the esterification reaction with fatty acids under lipase catalysis. The reaction efficiency was optimized using different poly-alcohols that were existed in the fermentation broth reacted with a fatty acid. Whereas the 1,3-PD conversion reached 62%, only a 0.06% and 0.08% conversion was reached for 2,3-butanediol and glycerol, illustrating the former's more efficient separation. The recovery efficiency of 1,3-PD was 96%. Finally, 1,3-PD was obtained by lipase-directed ester hydrolysis. Taken together, the bio-catalyzed separation process presented here is a novel and promising method for recovering 1,3-PD.

The impact of lncRNA MG3 on laryngeal cancer cell growth, cycle, and apoptosis related factors.

Laryngeal cancer is a common head and neck malignant tumor. Long non-coding RNA (lncRNA) is a kind of RNA at the length more than 200 nucleotides that cannot code protein. They widely exist in the human genome. Maternally expressed gene 3 (MG3) is a kind of lncRNA that is associated with various malignant tumors development. This study explores the influence of MG3 on laryngeal cancer Hep-2 cell growth, cycle, and apoptosis related factors. Hep2 cells transfected by MG3 were treated as experimental group, while untransfected Hep2 cells were treated as control. MG3 expression was tested by real time PCR. Cell proliferation was evaluated by MTT assay. Cell cycle was determined by flow cytometry. Bcl-2, Bax, and survivin protein levels were detected by Western blot. MG3 expression significantly increased in Hep2 cells compared with control (P < 0.05). Hep2 cell OD value and cell percentage in S phase were obviously declined, while cell apoptosis were markedly enhanced (P < 0.05). OD value and cell percentage in S phase apparently reduced in 12 h, 24 h, and 48 h from experimental group (P < 0.05). Bcl-2 and survivin protein downregulated, while Bax protein elevated in experimental group following time extension (P < 0.05). MG3 overexpression inhibited laryngeal cancer Hep2 cell proliferation and arrested cell cycle with time dependence, which may achieve by suppressing Bcl-2 and survivin protein, and facilitating Bax protein expression.

Genipin Cross-Linked Glucose Oxidase and Catalase Multi-enzyme for Gluconic Acid Synthesis.

In this work, glucose oxidase (GOD) and catalase (CAT) were used simultaneously to produce gluconic acid from glucose. In order to reduce the distance between the two enzymes, and therefore improve efficiency, GOD and CAT were cross-linked together using genipin. Improvements in gluconic acid production were due to quick removal of harmful intermediate hydrogen peroxide by CAT. GOD activity was significantly affected by the proportion of CAT in the system, with GOD activity in the cross-linked multi-enzyme (CLME) being 10 times higher than that in an un-cross-linked GOD/CAT mixture. The glucose conversion rate after 15 h using 15 % glucose was also 10 % higher using the CLME than was measured using a GOD/CAT mixture.

Effect of chemical pretreatments on corn stalk bagasse as immobilizing carrier of Clostridium acetobutylicum in the performance of a fermentation-pervaporation coupled system.

In this study, different pretreatment methods were evaluated for modified the corn stalk bagasse and further used the pretreated bagasse as immobilized carrier in acetone-butanol-ethanol fermentation process. Structural changes of the bagasses pretreated by different methods were analyzed by Fourier transform infrared, crystallinity index and scanning pictures by electron microscope. And the performances of batch fermentation using the corn stalk based carriers were evaluated. Results indicated that the highest ABE concentration of 23.86g/L was achieved using NaOH pretreated carrier in batch fermentation. Immobilized fermentation-pervaporation integration process was further carried out. The integration process showed long-term stability with 225-394g/L of ABE solvents on the permeate side of pervaporation membrane. This novel integration process was found to be an efficient method for biobutanol production.

Immobilization of Yarrowia lipolytica lipase Ylip2 for the biocatalytic synthesis of phytosterol ester in a water activity controlled reactor.

In this work, phytosterol ester was synthesized using Yarrowia lipolytica lipase Ylip2 that had been immobilized on inorganic support in a solvent-free system and reacted in a computer-aided water activity controlled bioreactor. The immobilization of Ylip2 on celite led to a remarkable increase in the phytosterol conversion compared to that of free lipase. An investigation of the reaction conditions were oleic acid as the fatty acid variety, 10,000U/g substrate, and a temperature of 50°C for phytosterol ester synthesis. Controlling of the water activity at a set point was accomplished by the introduction of dry air through the reaction medium at a digital feedback controlled flow rate. For the esterification of phytosterol ester, a low (15%) water activity resulted in a considerable improvement in phytosterol conversion (91.1%) as well as a decreased reaction time (78h). Furthermore, Ylip2 lipase immobilized on celite retained 90% esterification activity for the synthesis of phytosterol oleate after reused 8 cycles, while free lipase was only viable for 5 batches with 90% esterification activity remained. Finally, the phytosterol oleate space time yield increased from 1.65g/L/h with free lipase to 2.53g/L/h with immobilized lipase. These results illustrate that the immobilized Yarrowia lipolytica lipase Ylip2 in a water activity controlled reactor has great potential for the application in phytosterol esters synthesis.

Immobilizing Yarrowia lipolytica Lipase Lip2 via Improvement of Microspheres by Gelatin Modification.

The purpose of this study was to investigate the feasibility of immobilizing Yarrowia lipolytica lipase lip2 on epoxy microspheres with or without gelatin modifications. The activity of lipase immobilized on gelatin-modified supports was twofold higher than those immobilized on native supports. There was no significant difference in the Michaelis-Menten constant (K M ) between the two immobilized lipases. However, lipase immobilized on gelatin modified supports showed an approximately fourfold higher V max than lipase immobilized on native supports. Lipase immobilization on the gelatin-modified support exhibited a significantly improved operational stability in an esterification system. After it was reused for a total of 35 batches, the ester conversion of lipase immobilized on gelatin-modified and native microspheres was 83 and 60 %, respectively. Furthermore, the immobilized lipase could be stored at 4 °C for 12 months without any loss of activity.