Acid-induced conformation regulation of peanut polysaccharide and its effect on stability and digestibility of oil-in-water emulsion.

BACKGROUND: Developing the stable and healthy emulsion-based food is in accord with the needs of people for health. In the present study, acidification at pH 3.0 of peanut polysaccharide (APPSI) was employed to regulate its conformation and further improve its advantages in preparing oil-in-water emulsion. RESULTS: The results indicated that acidification induced conversion of PPSI aggregates into linear chains. Increasing concentration promoted formation of cross-linked network structure shown in transmission electron microscopy images. Consequently, the viscosity, yield stress, storage modulus and flow activation energy significantly increased, further fabricating gel structure. Moreover, aggregation behavior suggested that more exposed proteins were involved in gel structure, thereby forming many hydrophobic cores as verified by fluorescence spectroscopy of pyrene. Afterwards, emulsion characteristics indicated that APPSI produced strong and thick steric hindrance around oil droplets and the coil-like interweaved chains locked the continuous phase, bringing strong elasticity and resistance to stress and creaming. Meanwhile, the lower fatty acid in APPSI-emulsion was released after simulated gastrointestinal digestion, mainly as a result of the high retention ratio of emulsion droplets. Furthermore, the elastic and viscous Lissajous curves suggested that the structure strength of APPSI-emulsion was similar to that of the salad dressing within the strain 53.22%. CONCLUSION: The conformation of PPSI after acidification at pH 3.0 was suitable for preparing the stable emulsion. The obtained emulsion could resist digestion and maintain a strong structure, comprising a cholesterol-free and low-fat salad dressing substitute. © 2023 Society of Chemical Industry.

A clinical case report of Balamuthia granulomatous amoebic encephalitis in a non-immunocompromised patient and literature review.

BACKGROUND: Balamuthia granulomatous amoebic encephalitis (GAE) is a peculiar parasitic infectious disease of the central nervous system, about 39% of the infected Balamuthia GAE patients were found to be immunocompromised and is extremely rare clinically. The presence of trophozoites in diseased tissue is an important basis for pathological diagnosis of GAE. Balamuthia GAE is a rare and highly fatal infection for which there is no effective treatment plan in clinical practice. CASE PRESENTATION: This paper reports clinical data from a patient with Balamuthia GAE to improve physician understanding of the disease and diagnostic accuracy of imaging and reduce misdiagnosis. A 61-year-old male poultry farmer presented with moderate swelling pain in the right frontoparietal region without obvious inducement three weeks ago. Head computed tomography(CT) and magnetic resonance imaging(MRI) revealed a space-occupying lesion in the right frontal lobe. Intially clinical imaging diagnosed it as a high-grade astrocytoma. The pathological diagnosis of the lesion was inflammatory granulomatous lesions with extensive necrosis, suggesting amoeba infection. The pathogen detected by metagenomic next-generation sequencing (mNGS) is Balamuthia mandrillaris, the final pathological diagnosis was Balamuthia GAE. CONCLUSION: When a head MRI shows irregular or annular enhancement, clinicians should not blindly diagnose common diseases such as brain tumors. Although Balamuthia GAE accounts for only a small proportion of intracranial infections, it should be considered in the differential diagnosis.

Highly efficient isolation and purification of high-purity tea saponins from industrial camellia oil production by porous polymeric adsorbents.

BACKGROUND: Recovery of high-purity tea saponin (TS), a promising non-ionic surfactant with well-documented properties, is one of the major challenges to broadening its industrial applications. In this study, an innovative and sustainable strategy for the highly-efficient purification of TS was developed by using well-designed highly-porous polymeric adsorbents. RESULTS: The prepared Pp-A with controllable macropores (~96 nm) and appropriate surface hydrophobic properties was found more favorable for achieving high adsorption efficiency towards TS/TS-micelles. Kinetic results showed the adsorption follows the pseudo-second-order model (R2 = 0.9800), and the Langmuir model is more qualified to explicate the adsorption isotherms with Qe-TS ~ 675 mg g-1 . Thermodynamic studies revealed the monolayer adsorption of TS was an endothermic process that was conducted spontaneously. Interestingly, ethanol-driven desorption (90% v/v ethanol) of TS was rapidly (< 30 min) complete due to the possible ethanol-mediated disassembling of TS-micelles. A possible mechanism that involves the interactions between the adsorbents and TS/TS-micelles, the formation and disassembling of TS-micelles was proposed to account for the highly efficient purification of TS. Afterwards, Pp-A-based adsorption method was developed to purify TS directly from industrial camellia oil production. Through selective adsorption, pre-washing, and ethanol-driven desorption, the applied Pp-A enabled the direct isolation of high-purity TS (~96%) with a recovery ratio > 90%. Notably, Pp-A exhibited excellent operational stability and is of high potential for long-term industrial application. CONCLUSION: Results ensured the practical feasibility of the prepared porous adsorbents in purifying TS, and the proposed methodology is a promising industrial-scale purification strategy. © 2023 Society of Chemical Industry.

Radio frequency as an innovative method to produce low-fat French fries.

BACKGROUND: A large amount of evidence shows that excessive fat intake can increase the risk of obesity, type 2 diabetes, non-alcoholic fatty liver disease, and cardiovascular disease. The main purpose of this study was to use radio frequency (RF) technology to prepare low-fat French fries. RESULTS: RF treatment for 10 min significantly decreased the force required to cut potatoes and inhibited the enzymatic browning of fresh-cut potatoes. Moreover, RF treatment increased the hardness, gumminess, and chewiness of French fries from 388.55 g, 85.67, and 33.27 to 776.93 g, 159.36, and 70.11, respectively. Furthermore, RF treatment for 10 min reduced the oil content of French fries by 28.0% compared to that of the control group. This result was related to the pre-gelatinized potato starch content after RF treatment. Pre-gelatinized starch forms a 'protective film', that prevents oil from entering the fries during frying. CONCLUSION: Moderate RF treatment (10 min) reduced the oil content of French fries without making their texture significantly different from that of commercial French fries. These findings may provide a new perspective for the application of RF technology in the development of low-fat fried foods. © 2022 Society of Chemical Industry.

The application of the lytic domain of endolysin from Staphylococcus aureus bacteriophage in milk.

Staphylococcus aureus is a widespread foodborne pathogen that threatens human health. In particular, multidrug-resistant bacteria such as methicillin-resistant Staphylococcus aureus (MRSA) are emerging problems in modern health care, food safety, and animal health, which require the development of new antimicrobials to replace overused conventional antibiotics. Dairy products can potentially act as vehicles for the transmission of S. aureus and other antibiotic-resistant strains from the farm into the general human population, and should be controlled during the production and storage process. Recently, bacteriophage endolysins, which degrade the cell wall that is indispensable for bacteria, have been deemed promising antimicrobial agents. In this study, one endolysin, LysGH15, demonstrated prominent antimicrobial efficacy against S. aureus, as did its catalytic domain, cysteine, histidine-dependent amidohydrolase/peptidases (CHAP)LysGH15 alone. The LysGH15 and CHAPLysGH15 exhibited different characteristics in one MRSA strain (MRSA 2701), reaching the highest activity under different conditions (35°C and pH 6.0 for LysGH15, 40°C and pH 9.0 for CHAPLysGH15). A difference in the sensitivity of LysGH15 and CHAPLysGH15 to NaCl concentration was found, where the lytic activity of LysGH15 depends strongly on its binding domain's binding capacity, which is positively correlated with the NaCl concentration, whereas the CHAPLysGH15 activity showed a negative correlation with the NaCl concentration. When the NaCl concentration was 450 mM, the lytic activity of LysGH15 reached its peak, whereas the lytic activity of CHAPLysGH15 was the highest in the absence of NaCl. The difference in NaCl sensitivity between LysGH15 and CHAPLysGH15 may be due to the sensitivity of the SH3b binding protein of LysGH15 to NaCl. The CHAPLysGH15 was tested as a biopreservative in whole and skim milk and exerted effective control against S. aureus (declined by approximately 2.5 log10 cfu/mL when incubated at 4°C for 8 h), which suggests promise for application in dairy products.

Screening of a Bacillus subtilis strain producing both nattokinase and milk-clotting enzyme and its application in fermented milk with thrombolytic activity.

Bacillus subtilis is a generally recognized as safe probiotic, which is used as a starter for natto fermentation. Natto is a functional food with antithrombus function due to nattokinase. Compared with natto, fermented milk is a more popular fermented food, which is commonly fermented by Lactobacillus bulgaricus and Streptococcus. However, there is no report on B. subtilis-fermented milk. In this study, to produce a functional fermented milk with antithrombus function, a B. subtilis strain (B. subtilis JNFE0126) that produced both nattokinase and milk-clotting enzyme was isolated from traditionally fermented natto and used as the starter for the functional fermented milk. In liquid fermentation culture, the peak values of thrombolytic activity and milk-clotting activity were 3,511 U/mL at 96 h and 874.5 Soxhlet unit/mL at 60 h, respectively. The optimal pH and temperature were pH 7.0 at 40°C for nattokinase and pH 6.5 and 55°C for milk-clotting enzyme, respectively. The thrombolytic activity in the fermented milk reached 215.1 U/mL after 8 h of fermentation. Sensory evaluation showed that the acceptance of the milk fermented by B. subtilis JNFE0126 was similar to the traditional milk fermented by L. bulgaricus and S. thermophilus. More importantly, oral intake of the fermented milk by the thrombosis-model mice prevented the development of thrombosis. Our results suggest that B. subtilis JNFE0126-fermented milk has potential as a novel, functional food in the prevention of thrombosis-related cardiovascular diseases.

Preparation of high-quality sunflower seed protein with a new chlorogenic acid hydrolase from Aspergillus niger.

OBJECTIVE: To investigate the biochemical and enzymatic properties of chlorogenic acid hydrolase (ChlH) from Aspergillus niger SD14.721 and its applicability in sunflower seed protein processing. RESULTS: The ChlH with two identical subunits (97 kDa) was highly stable. Its optimal temperature and pH were determined as 60 °C and pH 7.0. The Km towards chlorogenic acid (CGA) was 1.85 µM. Based on its N-terminal sequence (AVDSVDAIFA), the purified ChlH appeared to be a new chlorogenic acid hydrolase. When applied in sunflower seed protein extraction, ChlH removed 99.13% of CGA in sunflower seed pastes, thus the colour of sunflower seed protein (SSP) changed from green to grey and its visual acceptance improved. Meanwhile, the solubility, water absorption capacity, and emulsification stability of SSP were increased 48.39%, 59.32% and 22.92%, respectively. CONCLUSIONS: A new ChlH was obtained and its feasibility as a CGA-removal tool to obtain high quality SSP was demonstrated.

Combination of thermal pretreatment and alcohol-assisted aqueous processing for rapeseed oil extraction.

BACKGROUND: The alcohol-assisted aqueous extraction processing (AAEP) of oil has many advantages such as no need for demulsification and relative low cost compared with enzymatic aqueous extraction processing (EAEP). Three kinds of thermal pretreatments including dry-heating, wet-heating and soak-heating followed by the AAEP of rapeseed oil were investigated. RESULTS: Both soak-heating and wet-heating had a higher contribution rate to oil yield than dry-heating due to the enhancement of heat transfer rate owing to the high moisture content in the rapeseed cells. However, oil from soak-heated rapeseeds showed a much lower level on peroxide value (0.41 mmol kg-1 ) than that of wet-heated rapeseeds (5.23 mmol kg-1 ). In addition, transmission electron microscopy images illustrated that promoting effects of soak-heating and wet-heating on oil release were attributed to the coalescence of oil bodies. A relative low concentration of alcohol solution as an extraction medium, the highest oil recovery of 92.77% was achieved when ground rapeseeds (mean particle size: 21.23 µm) were treated with 45% (v/v) alcohol for 2 h at 70 °C and pH 9.0. Both the acid value and the peroxide value are lower than the commercial oil produced by extrusion and hexane extraction. Furthermore, the oil produced from AAEP also had higher content of tocopherols and lower content of trans-fatty acids than the commercial oil. CONCLUSION: AAEP of oil from soak-heated rapeseeds is a promising alternative to conventional oil extraction methods. © 2019 Society of Chemical Industry.

Preparation of high-purity lactulose through efficient recycling of catalyst sodium aluminate and nanofiltration: a pilot-scale purification.

BACKGROUND: Lactulose, a valuable lactose-originated 'bifidus factor' product, is exclusively produced by chemical-based isomerization commercially. A complexing agent of sodium aluminate exhibiting high conversion efficiency and strong recyclable stability is more practical for industrial applications. In this study, efficient purification of high-purity lactulose through recycling of sodium aluminate and further desalination by nanofiltration (NF) was implemented on a pilot scale. RESULTS: Over 99.5% of the catalyst was prior recycled in the form of Al(OH)3 precipitate by pH-induced precipitation and centrifugation; residual aluminum was further absorbed by ion exchange resin to an acceptable level (≤10 mg kg-1 ). Subsequently, impurities (monosaccharides and NaCl) were ideally separated from lactulose syrup by NF based on their significant retention differences (lactulose 94.8-97.2% > lactose 86.2-93.5% > monosaccharides 36.3-48.7% > NaCl 9.5-31.1%). High-purity (>95%) lactulose was obtained with >90% yield in both constant and variable volume diafiltration (CVD and VVD) modes when the volume dilution ratio (Vc /Vf ) was 4.0 and 2.5 respectively. Both experimental and predicted results showed that the VVD mode was more water-saving in practice. CONCLUSION: This is the first trial purification of lactulose syrup from chemical isomerization of lactose catalyzed by sodium aluminate, and the applied methodology is a promising industrial-scale purification strategy. © 2018 Society of Chemical Industry.

Glycyrrhetic Acid 3-O-Mono-ß-d-glucuronide (GAMG): An Innovative High-Potency Sweetener with Improved Biological Activities.

Glycyrrhetic acid 3-O-mono-ß-d-glucuronide (GAMG) is an important derivative of glycyrrhizin (GL) and has attracted considerable attention, especially in the food and pharmaceutical industries, due to its natural high sweetness and strong biological activities. The biotransformation process is becoming an efficient route for GAMG production with the advantages of mild reaction conditions, environmentally friendly process, and high production efficiency. Recent studies showed that several ß-glucuronidases (ß-GUS) are key GAMG-producing enzymes, displaying a high potential to convert GL directly into the more valuable GAMG and providing new insights into the generation of high-value compounds. This review provides details of the structural properties, health benefits, and potential applications of GAMG. The progress in the development of the biotransformation processes and fermentation strategies to improve the yield of GAMG is also discussed. This work further summarizes recent advances in the enzymatic synthesis of GAMG using ß-GUS with emphasis on the physicochemical and biological properties, molecular modifications, and enzymatic strategies to improve ß-GUS biocatalytic efficiencies. This information contributes to a better framework to explore production and application of bioactive GAMG.

Recent progress in Bacillus subtilis spore-surface display: concept, progress, and future.

With the increased knowledge on spore structure and advances in biotechnology engineering, the newly developed spore-surface display system confers several inherent advantages over other microbial cell-surface display systems including enhanced stability and high safety. Bacillus subtilis is the most commonly used Bacillus species for spore-surface display. The expression of heterologous antigen or protein on the surface of B. subtilis spores has now been practiced for over a decade with noteworthy success. As an update and supplement to other previous reviews, we comprehensively summarize recent studies in the B. subtilis spore-surface display technique. We focus on its benefits as well as the critical factors affecting its display efficiency and offer suggestions for the future success of this field.

Characterization of a cold-adapted esterase and mutants from a psychotolerant Pseudomonas sp. strain.

A cold-adapted esterase-producing strain named T1-39 was isolated from Glacier No. 1, Tianshan, People's Republic of China and identified as Pseudomonas sp. from 16S rRNA sequence analysis. The esterase (EstT1-39) secreted by this strain preferentially hydrolyzed esters of glycerol with short- and medium-chain fatty acids. Mutants of T1-39 were generated by the atmospheric and room temperature plasma method and screened for enhanced esterase activity. Among all the mutants, strain TB11 had 4.45-fold higher esterase productivity than T1-39, with high genetic stability over 10 generations of continuous cultivation. Maximum activity of EstT1-39 and EstTB11 was observed at 30 ℃, pH 9.0 and 25 ℃, pH 8.5, respectively. EstTB11 was thermally more stable (50 ℃ for 1 H) and active over a broader pH range than EstT1-39. EstTB11 also retained 38% of its maximal activity at 0 ℃ and was found to be able to hydrolyze milk fats into short- and medium-chain fatty acids at 4 ℃. The characteristics of EstT1-39 made it a cold-adapted enzyme and the EstTB11 from the mutant, with its higher activity at lower temperatures, may be suitable for the production of aromas and flavors in the dairy industry.

miR-100 Inhibits the Growth and Migration of Burn-Denatured Fibroblasts.

BACKGROUND Burn-denatured dermis is able to regain the function and shape of normal dermis; however, the potential mechanisms are still vague. The aim of this study was to investigate roles of miR-100 involved in the growth and migration of burn-denatured fibroblasts. MATERIAL AND METHODS Quantitative real-time polymerase chain reaction(qRT-PCR) was used to assess the expression of miR-100. Transient transfection of miR-100 mimics and inhibitor was used to up-regulate or down-regulate the expression of miR-100. Cell proliferation and colony formation assay were used to test the cell growth, and wound healing assay and transwell migration assay were used to evaluate the cell migration. RESULTS miR-100 expression was notably downregulated in the burn-denatured fibroblasts compared to normal controls. Functionally, transfection of miR-100 inhibitors improved the growth and migration abilities of burn-denatured fibroblasts. In contrast, upregulation of miR-100 inhibits the growth and migration of burn-denatured fibroblasts. CONCLUSIONS Based on these observations, we concluded that miR-100 can inhibit the growth and migration of burn-denatured fibroblasts.

Aggregation of egg white proteins with pulsed electric fields and thermal processes.

BACKGROUND: Pulsed electric field (PEF) processing is progressing towards application for liquid egg to ensure microbial safety. However, it usually causes protein aggregation, and the mechanism is still unclear. In this study, egg white protein was applied to investigate the changes in protein structure and mechanism of aggregates formation and a comparison was made with thermal treatment. RESULTS: Soluble protein content decreased with the increase of turbidity after both treatments. Fluorescence intensity and free sulfhydryl content were increased after being treated at 70 °C for 4 min. Less-remarkable changes of hydrophobicity were observed after PEF treatments (30 kV cm(-1) , 800 µs). Soluble and insoluble aggregates were observed by thermal treatment, and disulfide bonds were the main binding forces. The main components of insoluble aggregates formed by thermal treatment were ovotransferrin (30.58%), lysozyme (18.47%) and ovalbumin (14.20%). While only insoluble aggregates were detected during PEF processes, which consists of ovotransferrin (11.86%), lysozyme (21.11%) and ovalbumin (31.07%). Electrostatic interaction played a very important role in the aggregates formation. CONCLUSION: PEF had a minor impact on the structure of egg white protein. PEF had insignificant influence on heat-sensitive protein, indicating that PEF has potential in processing food with high biological activity and heat sensitive properties. © 2015 Society of Chemical Industry.

Efficient and eco-friendly extraction of corn germ oil using aqueous ethanol solution assisted by steam explosion.

An improved aqueous extraction method has been established for extraction of oil from corn germs. This method primarily included steam explosion pretreatment and aqueous ethanol extraction. Process variables such as steam pressure, resident time, particle size and ethanol concentration were investigated. The highest yield of 93.74 % was obtained when ground steam-exploded corn germ (1.3 MPa, 30 s, 30-35 µm particle size) was treated with 30 % (v/v) aqueous ethanol for 2 h, at 60 °C and pH 9.0. The residual oil content in water and sediment phase decreased dramatically to 4 % and 3 %, respectively. The enhancement mechanism of the process induced by steam explosion was analyzed by confocal laser scanning microscope (CLSM). The quality of extracted crude oil was also investigated. The results showed that the quality of extracted oil was superior to commercial oils.

Cloning, expression and structural stability of a cold-adapted ß-galactosidase from Rahnella sp. R3.

A novel gene was isolated for the first time from a psychrophilic gram-negative bacterium Rahnella sp. R3. The gene encoded a cold-adapted ß-galactosidase (R-ß-Gal). Recombinant R-ß-Gal was expressed in Escherichia coli BL21 (DE3), purified and characterized. R-ß-gal belongs to the glycosyl hydrolase family 42. Circular dichroism spectrometry of the structural stability of R-ß-Gal with respect to temperature indicated that the secondary structures of the enzyme were stable to 45°C. In solution, the enzyme was a homo-trimer and was active at temperatures as low as 4°C. The enzyme did not require the presence of metal ions to be active, but Mg(2+), Mn(2+), and Ca(2+) enhanced its activity slightly, whereas Fe(3+), Zn(2+) and Al(3+) appeared to inactive it. The purified enzyme displayed K(m) values of 6.5 mM for ONPG and 2.2mM for lactose at 4°C. These values were lower than the corresponding K(m)s reported for other cold-adapted ß-Gals.

Adsorption-based immobilization of Caldicellulosiruptor saccharolyticus cellobiose 2-epimerase on Bacillus subtilis spores.

Nonrecombinant spore was examined as a novel immobilization support to adsorb enzymes. Caldicellulosiruptor saccharolyticus cellobiose 2-epimerase (CsCE), efficiently producing lactulose using lactose as a single substrate, was immobilized on Bacillus subtilis spores via adsorption. The immobilization process was optimized, and the properties of immobilized CsCE and the interactions between the enzyme and spores were also investigated. Under the optimized conditions (pH 4.5, temperature 4 °C, reaction time 2 H, and initial enzyme concentration 2.4 mg/mL), the maximum adsorbed amount of CsCE was 1.47 mg/10(11) spores, and the enzyme activity recovery was 79.4%. The spore-immobilized CsCE presented a higher pH and thermal stability than a free enzyme. Total desorption of the immobilized enzyme was only achieved by treatment with 1.0 M NaCl at pH 1.0, indicating a strong adsorption between CsCE and B. subtilis spores. Efficient binding may require a potent combination of electrostatic and hydrophobic interactions between spores and an enzyme. The immobilized CsCE was applied to produce 395 g/L lactulose after 4 H. Moreover, the spores could be regenerated and the spore-immobilized enzyme showed good reusability as it retained approximately 70% of its initial activity after eight recycles.

Extraction of rebaudioside-A by sonication from Stevia rebaudiana Bertoni leaf and decolorization of the extract by polymers.

Optimization of steviol glycosides extraction from Stevia rebaudiana Bertoni leaf was carried out by investigating the effects of isopropyl alcohol concentration (60 %, v/v), time (6-24 min), temperature (30 °C) and sonic power (300-480 W) on extraction of rebaudioside A from Stevia rebaudiana leaves and decolorization of the extract by polymer (Separan AP30 and Resin ADS-7). The results showed that isopropyl alcohol was suitable for the extraction of rebaudioside A from Stevia rebaudiana leaves and the yield of rebaudioside A achieved 35.61 g/100 g when the output power was 360 W and treatment time was 18 min. The sonication had influence on the particle size of stevia leaf and the color of the extracted solution. As the sonication intensity increased, the particle size decreased. The colour of differently treated stevia solutions were significantly different (P < 0.05). Separan AP30 and adsorption resin ADS-7 were performed to remove the colour impurity. The results showed that more than 65 % of the coloured impurity was removed by Separan AP30 combined with Calcium oxide (CaO).

Evaluation of the associations between vascular endothelial function and coronary artery stenosis in patients with elevated blood pressure during coronary angiography.

OBJECTIVES: The aim of the present study is to explore the correlation between vascular endothelial function and coronary artery stenosis in non-hypertensive patients with elevated blood pressure under stress. METHODS: This study included 1141 patients suspected of having coronary artery disease (CAD) without hypertension. Coronary arteriography and ultrasonic detection were used to measure the flow-mediated dilatation (FMD) function in the brachial artery. Patients were divided into 2 groups according to coronary angiography: experiment group, patients with blood pressure ≥ 140/90 mm Hg; control group, patients with blood pressure <140/90 mm Hg. The correlation between vascular endothelial function and coronary artery stenosis was observed. RESULTS: The majority of the patients in the control group were found to have either normal coronary arteries or stenosis <50%. Patients in the experiment group (those with invasive blood pressure [IBP] >140/90) were more likely to have some degree of coronary artery stenosis. Specifically, there were significantly more patients with >50% stenosis in the experiment when compared with the control group (P < .05). The FMD in the experiment group was significantly lower than that in the control group (P < .05). CONCLUSION: The non-hypertensive patients with elevated blood pressure under stress had coronary artery stenosis, which was associated with vascular endothelial dysfunction.

Expression and characterization of two ß-galactosidases from Klebsiella pneumoniae 285 in Escherichia coli and their application in the enzymatic synthesis of lactulose and 1-lactulose.

The two genes lacZ1 and lacZ2 from Klebsiella pneumoniae 285, encoding ß-galactosidase isoenzymes II and III (KpBGase-II and -III), were each cloned downstream of a T7 promoter for expression in Escherichia coli BL21(DE3), and the resulting recombinant enzymes were characterized in detail. The optimum temperature and pH value of KpBGase-II were 40 °C and 7.5, and those of KpBGase-III were 50 °C and 8.0, respectively. KpBGase-III was more stable than KpBGase-II at higher temperature (>60°C). Both ß-galactosidases were more active towards o-nitrophenyl-ß- D-galactopyranoside as compared to lactose. The enzymatic synthesis of lactulose and 1-lactulose catalyzed by KpBGase-II and KpBGase-III was investigated. Using 400 g/L lactose and 200 g/L fructose as substrates, the resulting lactulose and 1-lactulose yields with KpBGase-II were 6.2 and 42.3 g/L, while those with KpBGase-III were 5.1 and 23.8 g/L, respectively. KpBGase-II has a potential for the production of 1-lactulose from lactose and fructose. Like other ß-galactosidases, the two isozymes catalyze the transgalactosylation in the presence of fructose establishing the ß-(1→1) linkage.