Foods
·
2025
·Vol. 14
·期 11
·1873
·
IF 5.1
Zhou Chen,Yuhan Yan,Ziang Wu,Yanyin Song,Jiangqi Xu
As global milk production continues to rise, the disposal of expired milk contributes to environmental pollution and valuable resource wastage. This study presents the development of a novel L-arabinose isomerase, designatedBmAIase12, and its application in the enzymatic recycling of expired milk.BmAIase12 exhibited a specific activity of 10.7 U/mg and showed optimal performance at 50 °C and pH 7.0. Furthermore, it exhibited higher activity than most other L-arabinose isomerases. It converted D-galactose into D-tagatose with a high conversion ratio of 53.3% after 48 h at 50 °C. The conversion efficiency of expired milk to D-tagatose was recorded at 40.62%, resulting in a maximum tagatose yield of 1.625 g/L. This was accomplished through the incorporation of β-galactosidase (120 U/mL) andSaccharomyces cerevisiae(30 mg/mL) to hydrolyze lactose and metabolize glucose, followed by the addition of 3 U/mL ofBmAIase12. Ultimately, following purification, the purity of tagatose was determined to be 98%, and the final yield was 29.8%. These results suggest thatBmAIase12 may serve as a promising enzyme for D-tagatose production due to its high conversion yield.
研究领域:
生物化学
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环境科学
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食品科学
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产品关联
DOI
PubMed
高影响因子
Foods
·
2025
·Vol. 14
·期 9
·1538
·
IF 5.1
Zepeng Li,Runmin Wang,Xiantai Lai,Wenyi Liao,Runfeng Liao,Zhuohong Wu,Guoyan Zhang,Xianghui Qi
D-tagatose is a functional sweetener with glucose-regulating and prebiotic properties, but its bioproduction from D-galactose faces many limitations, particularly the high production costs. In particular, the current biosynthesis of D-tagatose suffers from thermal instability and the substrate selectivity issues of L-arabinose isomerase (L-AI) required to convert D-galactose into D-tagatose. In this study, recombinantEscherichia coliBW25113/pQE-80L-araAF118M/F279Iexpressing double mutant L-AI was immobilized to enhance its stability and reusability. The optimal conditions for whole-cell catalysis were 60 °C, pH 6.5, 5 mM Mn2+, and 20 h, with a yield of 55.2 g/L of D-tagatose. Immobilization with 3% sodium alginate and 2% CaCl2retained 90% of the production efficiency displayed by free cells. Notably, the immobilized cells exhibited enhanced heat resistance (60–70 °C) and operational stability, retaining 76% activity after five cycles. The D-tagatose production was further increased to 129.43 g/L by increasing the substrate concentration to 250 g/L. Compared to free cells, immobilized cells retained 83.6% of the initial yield up to 10 batches. This study presents a cost-effective and sustainable method for the production of D-tagatose using optimized whole-cell catalysis through immobilization, which paves the way to solve industrial challenges such as thermal instability and low substrate efficiency.
研究领域:
生物化学
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食品科学
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产品关联
DOI
PubMed
高影响因子
Foods
·
2024
·Vol. 13
·期 11
·1727
·
IF 4.7
Chengyu Lu,Ziwei Chen,Yuvaraj Ravikumar,Guoyan Zhang,Xinrui Tang,Yufei Zhang,Mei Zhao,Wenjing Sun,Xianghui Qi
L-Arabinose isomerase (L-AI) has been commonly used as an efficient biocatalyst to produce D-tagatose via the isomerization of D-galactose. However, it remains a significant challenge to efficiently synthesize D-tagatose using the native (wild type) L-AI at an industrial scale. Hence, it is extremely urgent to redesign L-AI to improve its catalytic efficiency towards D-galactose, and herein a structure-based molecular modification ofLactobacillus plantarumCY6 L-AI (LpAI) was performed. Among the engineered LpAI, both F118M and F279I mutants showed an increased D-galactose isomerization activity. Particularly, the specific activity of double mutant F118M/F279I towards D-galactose was increased by 210.1% compared to that of the wild type LpAI (WT). Besides the catalytic activity, the substrate preference of F118M/F279I was also largely changed from L-arabinose to D-galactose. In the enzymatic production of D-tagatose, the yield and conversion ratio of F118M/F279I were increased by 81.2% and 79.6%, respectively, compared to that of WT. Furthermore, the D-tagatose production of whole cells expressing F118M/F279I displayed about 2-fold higher than that of WT cell. These results revealed that the designed site-directed mutagenesis is useful for improving the catalytic efficiency of LpAI towards D-galactose.
研究领域:
生物化学
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食品科学
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分子生物学
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DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2014
·
IF 3.107
Yi Liu,Sha Li,Hong Xu,Lingtian Wu,Zheng Xu,Jing Liu,Xiaohai Feng
d-Tagatose, a functional sweetener, is commonly transformed from d-galactose by l-arabinose isomerase (l-AI). In this study, a novel type of biocatalyst, l-AI from Lactobacillus fermentum CGMCC2921 displayed on the spore surface of Bacillus subtilis 168, was developed for producing d-tagatose. The anchored l-AI, exhibiting the relatively high bioactivity, suggested that the surface display system using CotX as the anchoring protein was successfully constructed. The stability of the anchored l-AI was significantly improved. Specifically, the consolidation of thermal stability representing 87% of relative activity was retained even at 80 °C for 30 min, which remarkably favored the production of d-tagatose. Under the optimal conditions, the robust spores can convert 75% d-galactose (100 g/L) into d-tagatose after 24 h, and the conversion rate remained at 56% at the third cycle. Therefore, this biocatalysis system, which could express the target enzyme on the food-grade vector, was an alternative method for the value-added production of d-tagatose.
研究领域:
生物工程
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食品科学
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DOI
PubMed
高影响因子
CHINESE JOURNAL OF CHEMICAL ENGINEERING
·
2017
·
IF 1.174
Lei Guo,Yangdong Hu,Lianying Wu,Chen Liang,Weitao Zhang
To achieve green hydrolysis technology of hemicellulose through repeated using hydrolysate, the hydrolysis of hemicellulose in corncob was studied. The influence of repeated use of corncob hydrolysate on concentrations of d -xylose and l -arabinose was investigated. The loss rates of d -xylose in the prepared d -xylose solutions both with and without corncob, and repeated using corncob hydrolysate under identical acidity condition were discussed. The result shows that d -xylose concentration and l -arabinose concentration are all gradually increasing with the growing time of repeated use of corncob hydrolysate. After the fifth repetition, the concentrations of d -xylose and l -arabinose are 196.7 g·L − 1 and 22.0 g·L − 1 , respectively. Substance inhibiting the degradation of d -xylose is generated during repeated use of corncob hydrolysate, and the production is further proved by the change of d -xylose concentration and the loss rate of d -xylose over heating time.
研究领域:
生物技术
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化学工程
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可持续化学
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DOI
高影响因子
FOOD HYDROCOLLOIDS
·
2025
·
IF 12.4
Yan Wen,Yan Zhang,Na Lü,Zehui Xuan,Lili Lu
D-tagatose is an attractive functional sweetener, and its production via enzymatic synthesis is simple and environmentally friendly; however, it faces challenges related to enzyme and substrate costs. In this study, a novel and cost-effective method for synthesizing D-tagatose was established using L-arabinose isomerase (L-AI) immobilized within a sodium alginate (SA) composite hydrogel, with the low-cost guar gum as the starting material. Eight inorganic or organic materials were individually combined with SA to encapsulate L-AI. The immobilized enzyme prepared with SA and 50-nm SiO 2 particles demonstrated superior properties, including enhanced mechanical strength and improved reusability, compared to enzymes immobilized with other carriers. The structure of the immobilized enzyme was characterized by scanning electron microscopy, Fourier-transform infrared spectroscopy, and X-ray diffraction. The D-tagatose synthesis reaction using the immobilized enzyme was optimized through both single-factor and statistical methods. The maximum yield of D-tagatose reached 30.12% from 1 M D-galactose at 68°C, with a productivity of 6.78 g/L/h. The immobilized enzyme was reusable for 20 cycles of reaction at 60°C, retaining over 55% of its initial activity and accumulating a high amount of 863.74 g/L D-tagatose. Furthermore, it efficiently converted guar gum hydrolysate into D-tagatose, retaining over 50% enzyme activity and yielding 114.32 g/L of D-tagatose after 10 cycles. The excellent enzyme reusability and the exploration of an economic substrate would significantly reduce production costs in large-scale D-tagatose synthesis in the future.
研究领域:
生物化学
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食品科学
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产品关联
DOI
高影响因子
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
·
2020
·Vol. 192
·期 3
·935-951
·
IF 2.277
Guo Zongren,Long Liangkun,Ding Shaojun
L-Ribulose and L-ribose are two high-value unnatural sugars that can be biosynthesized by sugar isomerases. In this paper, an L-arabinose isomerase (BvAI) from Bacillus velezensis CICC 24777 was cloned and overexpressed in Escherichia coli BL21 (DE3) strain. The maximum activity of recombinant BvAI was observed at 45 °C and pH 8.0, in the presence of 1.0 mM Mn 2+ . Approximately 207.2 g/L L-ribulose was obtained from 300 g/L L-arabinose in 1.5 h by E. coli harboring BvAI. In addition, approximately 74.25 g/L L-ribose was produced from 300 g/L L-arabinose in 7 h by E. coli co-expressing BvAI and L-RI from Actinotalea fermentans ATCC 43279 (AfRI). This study provides a feasible approach for producing L-ribose from L-arabinose using a co-expression system harboring L-Al and L-RI.
研究领域:
生物技术
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生物化学
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微生物学
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产品关联
DOI
PubMed
高影响因子
BIORESOURCE TECHNOLOGY
·
2020
·
IF 7.539
Guoyan Zhang,Hossain M. Zabed,Junhua Yun,Jiao Yuan,Yufei Zhang,Yang Wang,Xianghui Qi
In this study, a new strain of Lactobacillus plantarum (CY.6) was identified and its L-arabinose isomerase (L-AI) encoding gene ( ara A) was overexpressed in Escherichia coli BL21 for the biosynthesis of D-tagatose from milk whey powders (WP). Whole-cell biotransformation of lactose in WP into D-tagatose was done by three technological approaches, including 100%, 50% and 0% hydrolysis of lactose in WP before biotransformation, where simultaneous saccharification and biotransformation (SSB, 0% prior hydrolysis of lactose) produced maximum amounts of D-tagatose. Two-stage SSB provided 73.6% conversion efficiency (based on D-galactose) and 36.8% (in term of lactose), with 51.5 g/L of D-tagatose after 96 h, while concentration of D-tagatose produced after first stage was 34.4 g/L. Yield and volumetric productivity of D-tagatose after two-stage SSB were found to be 0.26 g/g of WP (0.37 g/g of lactose, 0.74 g/g of D-galactose produced from lactose) and 0.54 g/L/h, respectively.
研究领域:
生物技术
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微生物学
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食品工程
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酶学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF NUTRITION
·
2023
·
IF 4.2
Luyuan Kang,Jiaman Pang,Xiangyu Zhang,Yisi Liu,Yujun Wu,Junjun Wang,Dandan Han
Background L-arabinose has anti-inflammatory and metabolism-promoting properties, and macrophages participate in the alleviation of inflammation; however, the mechanism by which they contribute to the anti-inflammatory effects of L-arabinose is unknown. Objectives To investigate the involvement of macrophages in the mitigation of L-arabinose in an intestinal inflammation model induced by lipopolysaccharide (LPS). Methods Five-week-old male C57BL/6 mice were divided into 3 groups: a control and an LPS group that both received normal water supplementation, and an L-arabinose (ARA+LPS) group that received 5% L-arabinose supplementation. Mice in the LPS and ARA+LPS groups were intraperitoneally injected with LPS (10 mg/kg body weight), whereas the control group was intraperitoneally injected with the same volume of saline. Intestinal morphology, cytokines, tight junction proteins, macrophage phenotypes, and microbial communities were profiled at 6 h postinjection. Results L-arabinose alleviated LPS-induced damage to intestinal morphology. L-arabinose down-regulated serum tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-6, and messenger RNA (mRNA) levels of TNF-α , IL-1β , interferon-γ ( IFN-γ ), and toll-like receptor-4 in jejunum and colon compared with those of the LPS group ( P < 0.05). The mRNA and protein levels of occludin and claudin-1 were significantly increased by L-arabinose ( P < 0.05). Interferon regulatory factor-5 ( IRF-5 ) and signal transducer and activator of transcription-1 ( STAT-1 ), key genes characterized by M1 macrophages, were elevated in the jejunum and colon of LPS mice ( P < 0.05) but decreased in the ARA+LPS mice ( P < 0.05). In vitro , L-arabinose decreased the proportion of M1 macrophages and inhibited mRNA levels of TNF-α , IL-1β , IL-6 , IFN-γ , as well as IRF-5 and STAT-1 ( P < 0.01). Moreover, L-arabinose restored the abundance of norank_ f__Muribaculaceae , Faecalibaculum , Dubosiella , Prevotellaceae_UCG-001 , and Paraasutterella compared with those of LPS ( P < 0.05) and increased the concentration of short-chain fatty acids ( P < 0.05). Conclusion The anti-inflammatory effects of L-arabinose are achieved by reducing M1 macrophage polarization, suggesting that L-arabinose could be a candidate functional food or nutritional strategy for intestinal inflammation and injury.
研究领域:
免疫学
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胃肠病学
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营养学
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微生物学
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2025
·
IF 8.5
Yun Liu,Zhina Qiao,Hengwei Zhang,Xian Zhang,Yanan Li,Zhiming Rao
D-tagatose, a functional rare sugar, has physiological effects such as lowering blood sugar and anti-caries. However, research on the industrial production of D-tagatose has primarily focused on Escherichia coli , limiting application in the food field. In this study, a cell factory for efficient synthesis of D-tagatose was constructed using the GRAS strain Corynebacterium glutamicum . Firstly, the key enzyme L-arabinose isomerase from Lactobacillus fermentum was screened. Secondly, by rational design, the optimal mutant D390V/H339P significantly enhanced enzyme activity by 40.65 % and improved thermostability. Subsequently, a dual-enzyme coupled whole-cell system was established by regulating the ratio of β-galactosidase and L-arabinose isomerase by RBS optimization. After optimizing the catalytic system, 101.42 g/L D-tagatose was produced from 400 g/L lactose, and the conversion rate reached 25.12 % in 5 L fermenter, which is the highest yield using lactose as substrate and GRAS strain as chassis cells. This research laid a foundation for the industrial application of food-grade D-tagatose.
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PubMed
高影响因子
Gels
·
2024
·Vol. 10
·期 12
·808
·
IF 5
Feng Li,Ruoxi Yang,Jian Xu,Guohai Xu,Ye Wu
The concentration of small molecules reflects the normality of physiological processes in the human body, making the development of simple and efficient detection equipment essential. In this work, inspired by a facile strategy in point-of-care detection, two devices were fabricated to detect small molecules via photocurrent measurement. A linear response of the photocurrent against the concentration of the small molecules was found. In the first device, metal ions were introduced into gel substrates made by xanthan gum to enhance photocurrent response. N-phenyl-2-naphthylamine was measured when iron or manganese ions were used. L-Phenylalanine was measured when the gel was modified by samarium, iron, cerium, or ytterbium ions. L-(+)-Arabinose was detected via the gels modified by iron or holmium ions. D-(+)-Mannose was detected when the gel was modified by ytterbium, manganese, chromium, or sodium ions. L-Methionine was detected in the gels modified by samarium, zinc, or chromium ions. The second device was based on a paper sheet. A sugar-like molecule was first synthesized, which was then used to modify the paper. The detection was possible since the photocurrent showed a linear trend against the concentration of D-Trehalose. A linear fit was conducted to derive the sensitivity, whose value was found to be 5542.4. This work offers a novel, simple, and environmentally sustainable platform that is potentially useful in remote areas lacking medical devices.
研究领域:
生物医学工程
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分析化学
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环境科学
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产品关联
DOI
PubMed
高影响因子
APPLIED BIOCHEMISTRY AND MICROBIOLOGY
·
2024
·
IF 1
Hu J.,Wu Y.,Kang L.,Liu Y.,Wu Y.,Han D.,Wang J.
Abstract Shiga toxin-producing Escherichia coli (STEC) is an important foodborne pathogen that seriously threats the public and animal health. The major virulence of STEC is Shiga toxin (Stx) which is encoded by Stx prophage. Stx prophage induction leads to the production of Stx. Our recent research has shown that dietary functional sugars can differently regulate Shiga toxin type 2 (Stx2) prophage induction. However, high-throughput assays are required to find more dietary inhibitors for Stx prophage induction. This study aims to establish the recA dual-luciferase reporter system for rapidly screening the functional sugars that can inhibit Stx prophage induction. Results showed that the recA dual-luciferase reporter plasmid was successfully constructed as confirmed by DNA sequencing. Established recA dual-luciferase reporter system showed similar screening results when compared with traditional temperate phage determination method (i.e., qPCR) and suggested that L-arabinose is a dietary functional Stx prophage induction inhibitor. Supplementation with L-arabinose in culture medium reduces Stx production in STEC isolated from fecal samples of diarrhea piglets. Collectively, the recA dual-luciferase reporter system is a promising tool for convenient and rapid screening of dietary Stx prophage induction inhibitor, especially suitable for large-scale screening experiments.
研究领域:
微生物学
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分子生物学
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食品安全
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生物技术
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）