Enhancing lipase-catalyzed production of erythorbyl ricinoleate: A strategy for water removal using adsorbents in a solvent-free bioreactor system Inwoo Park, Hyunjong Yu, Pahn-Shick Chang Lwt, 2025 This study aimed to enhance erythorbyl ricinoleate (ERO) production in a solvent-free bioreactor system by removing the water produced during lipase-catalyzed esterification. Reaction conditions were optimized with gas flow rate of 2.0 L/min, substrate molar ratio [(ricinoleic acid):(erythorbic acid)] of 2.0, and temperature at 60 °C. The initial water content was controlled between 0.015 g/100 g and 0.055 g/100 g with the highest initial reaction rate (0.020 ± 0.002 mmol/h) observed at a water content of 0.025 g/100 g. Removal of the water generated during esterification by adsorbents significantly increased the initial reaction rate, but excessive removal of water decreased the reaction rate. The initial reaction rate (0.034 ± 0.002 mmol/h) achieved by adding 10 mg/mL molecular sieve 4 Å at the beginning of the reaction was 1.65-fold higher than that without adsorbent (0.027 ± 0.001 mmol/h). These findings provide significant insights for optimizing the immobilized lipase-catalyzed production of organic compounds, particularly in solvent-free bioreactor systems. • Lipase-catalyzed esterification in solvent-free bioreactor system. • Various adsorbents selection to control water produced during esterification. • Determination of adsorbent addition time for effective water control to enhance production. • Verifying effect of adsorbent for water control in scaled-up reactor.
Cysteine protease I29 propeptide from Calotropis procera R. Br. As a potent cathepsin L inhibitor and its suppressive activity in breast cancer metastasis Yong-Jin Kwon, Juno Lee, Eun-Bi Seo, Juchan Lee, Jaehyeon Park, Seul-Ki Kim, Hyunjong Yu, Sang-Kyu Ye, Pahn-Shick Chang Scientific Reports, 2024 Breast cancer metastasis is associated with a poor prognosis and a high rate of mortality. Cathepsin L (CTSL) is a lysosomal cysteine protease that promotes tumor metastasis by degrading the extracellular matrix. Gene set enrichment analysis revealed that CTSL expression was higher in tumorous than in non-tumorous tissues of breast cancer patients and that high-level CTSL expression correlated positively with the epithelial-mesenchymal transition. Therefore, we hypothesized that inhibiting CTSL activity in tumor cells would prevent metastasis. In this study, we characterized the inhibitory activity of SnuCalCpI15, the I29 domain of a CTSL-like cysteine protease from Calotropis procera R. Br., and revealed that the propeptide stereoselectively inhibited CTSL in a reversible slow-binding manner, with an inhibitory constant ( K i ) value of 1.38 ± 0.71 nM, indicating its potency as an exogenous inhibitor in anti-cancer therapy. SnuCalCpI15 was localized intracellularly in MDA-MB-231 breast cancer cells and suppressed tumor cell migration and invasion. These results demonstrate the potential of SnuCalCpI15 as a novel agent to prevent breast cancer metastasis.
Lipase-catalyzed synthesis of erythorbyl oleate and its characterization as a multifunctional emulsifier Jihoon Kim, Yoonseok Choi, Hyunjong Yu, Pahn‐Shick Chang Journal of Food Science, 2024 Erythorbyl oleate (EO), a novel emulsifier with multifunctional properties, was synthesized via lipase‐catalyzed solvent‐free esterification between erythorbic acid and oleic acid. The reaction produced EO of 30.75 ± 1.65 mM at 60 h, monitored by high‐performance liquid chromatography (HPLC) analysis. Purified EO was chemically identified as 6‐O‐oleoyl‐erythorbic acid by HPLC‐electrospray ionization/mass spectrometry (ESI‐MS), 1H nuclear magnetic resonance (NMR), and 13C NMR analyses, and further investigations on its multifunctionalities were conducted. EO displayed antioxidant activity equivalent to erythorbic acid in 2,2‐diphenyl‐1‐picrylhydrazyl and 2,2′‐azino‐bis(3‐ethylbenzothiazoline‐6‐sulfonic acid) radical scavenging activities. Investigation of the antibacterial activity of EO against foodborne pathogens showed MICs of 0.27 ± 0.09 and 0.40 mM for Bacillus cereus ATCC 10876 and Staphylococcus aureus ATCC 12692, respectively. EO also displayed interfacial properties, lowering interfacial tension between medium‐chain triglyceride oil and water to 3.40 ± 0.04 mN/m with a critical micelle concentration of 2.54 ± 0.03 mM. Dynamic light scattering analyses of the emulsion stabilized by EO demonstrated a similar polydispersity index (0.15 ± 0.00) to Tween 80‐stabilized emulsion (0.28 ± 0.01), with an average droplet size of 173.2 ± 1.8 nm. In addition, the EO‐stabilized emulsion exhibited a negative surface charge (−56.83 ± 4.30 mV) in zeta potential measurements, indicating sufficient electrostatic repulsion between droplets. These results suggest that EO is a multifunctional emulsifier to simultaneously control oxidative rancidification and bacterial spoilage in emulsion‐based foods.Practical Application: EO was synthesized via an enzymatic esterification reaction between erythorbic acid and oleic acid. The synthetic process excluded any use of organic solvent, diminishing potential health hazards or environmental pollution associated with chemical reactions. The interfacial properties of the resultant EO, along with excellent antioxidant and antibacterial activities, propose its potential application in emulsion‐based foods. Simultaneous control of bacterial spoilage and oxidative rancidification by EO would fulfill the industrial needs for extending shelf life and enhancing consumer safety.
Bromelain-Decorated Nanoscale Liposomes for Mucus Permeation and Intestinal Absorption in Oral Drug Delivery Haena Park, Sunghak Choi, Bong Su Kang, Hyunjong Yu, Jaeil Kim, Ho-Sup Jung, Hoon Eui Jeong, Pahn-Shick Chang ACS Applied Nano Materials, 2024 Nanoscale liposomes are widely recognized as promising drug delivery materials; yet, their utilization in oral drug delivery has been hindered by their limited ability to efficiently penetrate the intestinal mucus layer. Herein, we present an approach to enhance oral drug delivery and improve the mucus penetration capability of liposomes by conjugating bromelain, a mucolytic enzyme derived from pineapple stems, to the liposome membrane (Bro-Lip). With an optimized bromelain payload of 11.1 nmol/mg, we achieved an impressive 47.8% conjugation efficiency while retaining 77.5% of the proteolytic activity post conjugation. Physicochemical analysis revealed that Bro-Lip measured 152.8 nm in size with a ζ potential of −2.3 mV. Notably, Bro-Lip exhibited a significantly enhanced capacity to penetrate the porcine mucus layer, demonstrating a 2.87-fold improvement compared with bare liposomes. Furthermore, we assessed the transcellular permeability of Bro-Lips using Caco-2 and Caco-2/HT29 coculture models, revealing substantial enhancements in permeability compared to unmodified liposomes. Specifically, in Caco-2 cell monolayers, mucus permeability increased by 3.80-fold, while in Caco-2/HT29 cocultures, it rose by 2.25-fold. Additionally, Bro-Lip demonstrated an improved cellular uptake in Caco-2 cells. These findings highlight the potential of Bro-Lip as an effective oral delivery system by enhancing mucus permeation and cell membrane permeation.
