Nine different isoenzymes and (or) isoforms of alkaline phosphatase (ALP; EC 3.1.3.1) from human tissue were studied with respect to Km and Vmax values for p-nitrophenyl phosphate (p-NPP) in seven different potential phosphoacceptors/buffers. Generally, the phosphoacceptors/buffers with the lowest affinity for p-NPP (highest Km values) gave the highest Vmax values; for the nine enzyme forms in this study, the mean Km and Vmax values were greatest in 2-(ethylamino) (EAE). The two amino-propanol buffers gave the lowest Km and Vmax values. The phosphoacceptors/buffers N-methyl-D-glucamine (MEG), diethanolamine, and Tris had intermediate Km and Vmax values. Hydrophilic liver ALP retained > 90% of its activity after 24 h at 30 degrees C in both 1.0 and 0.3 mol/L Tris and 2-amino-2-methyl-1,3-propanediol and in 0.3 mol/L MEG. This isoenzyme showed greatest inactivation upon prolonged exposure to 1.0 and 0.3 mol/L EAE, the activity at 24 h being approximately 50-66% of that at zero time. p-NPP underwent the greatest spontaneous degradation, approximately 2.5 times that of baseline levels, in 1 mol/L MEG. There was little degradation in all of the buffers tested at 0.3 mol/L or in Tris, EAE, and 2-amino-2-methyl-1-propanol at 1.0 mol/L.
Download full-text PDF |
Source |
|---|
Publication Analysis
Top Keywords
Similar Publications
Evolved enzymes in the metabolism of biological poly-acids: Applications in otolaryngological biocatalysis.
Int J Biol Macromol
January 2025
Department of Otolaryngology, The First Hospital of China Medical University, 155 Nanjing Street, Heping, Shenyang, Liaoning 110001, China. Electronic address:
This study explores evolved Hyaluronidase, Lipase, and Elastase's identification, characterization, and therapeutic potential to enhance tissue regeneration and drug delivery systems in otolaryngology. Hyaluronidase variant H5 exhibited a turnover number (k_cat) of 1500 min, a 200 % increase over wild-type (500 min), demonstrating superior hyaluronic acid degradation. Similarly, lipase variant L2 reached 1200 min (400 min wild-type), and elastase variant E3 showed a turnover of 2200 min (1000 min wild-type).
View Article and Find Full Text PDFDiscovery and characterization of a new β-glucosidase from Wolfiporia cocos through RNA sequencing and heterologous expression in Escherichia coli.
Int J Biol Macromol
January 2025
The Key Laboratory of Regenerative Biology, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.
De novo RNA-sequencing of Wolfiporia cocos mycelia cultured with filter paper composed of cellulose as the sole carbon source revealed a total of five expressed β-glucosidase genes. Among these, the β-glucosidase named Wcbg1B-1, which is composed of 539 amino acid residues and belongs to the GH1 family, had the highest mRNA abundance, accounting for 65 % of the total mRNA of the five expressed β-glucosidases. The recombinant Wcbg1B-1 was successfully expressed in Escherichia coli, with an optimal pH of 6.
View Article and Find Full Text PDFSynthesis of chiral alcohol (S)-CHBE by co-immobilization of double enzymes based on organic-inorganic hybrid nanoflower.
Int J Biol Macromol
January 2025
Key Laboratory of Geriatric Nutrition and Health, Ministry of Education, Beijing Technology and Business University (BTBU), Beijing 100048, China; School of Light Industry Science and Engineering, Beijing Technology and Business University (BTBU), Beijing 100048, China. Electronic address:
The chiral alcohols (S)-4-chloro-3-hydroxy-butyric acid ethyl ester ((S)-CHBE) is a critical intermediate in the synthesis of various active pharmaceutical ingredients. This study presents the first investigation of the efficient production of (S)-CHBE using organic-inorganic hybrid nanoflowers (GDH-CR@HNFs) for the co-immobilization of glucose dehydrogenase (BsGDH) and carbonyl reductase (BsCR). By optimizing immobilization conditions, we significantly enhanced the catalytic activity and immobilization efficiency of the hybrid nanoflowers.
View Article and Find Full Text PDFHydrolyzing collagen by extracellular protease Hap of Aeromonas salmonicida: Turning chicken by-products into bioactive resources.
Food Chem
January 2025
State Key Laboratory of Meat Quality Control and Cultured Meat Development, College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, PR China. Electronic address:
Collagen-rich meat processing by-products have potential utilization value. Extracellular protease Hap from meat-borne Aeromonas salmonicida has been identified as an ideal protease for hydrolyzing collagen. Here, to explore the possible application of Hap for giving chicken by-products a high added value, the hydrolysis ability and mechanism were investigated.
View Article and Find Full Text PDFImmobilization and characterization of β-galactosidase from Aspergillus oryzae in PVA-CMC hydrogel.
Int J Biol Macromol
January 2025
Department of Chemistry, Faculty of Arts and Science, Yildiz Technical University, Turkey. Electronic address:
Creating new formulations of immobilized enzymes has been a major focus of modern biotechnology. In this study, the industrially significant β-galactosidase was immobilized by being trapped in a polyvinyl alcohol and carboxymethyl cellulose (PVA-CMC) gel. The immobilized enzyme was optimized and characterized, and the results were compared with those obtained using free enzymes.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!