Rate Of Hydrolysis Research Articles

Hydrolysis rate and mechanism of water-dispersed polyesters with different sulfonate group contents in the cutinase-catalyzed system.

Water-dispersed polyester (WPET) has irreplaceable and wide application prospects in the textile field, but it is easy to form printing and dyeing wastewater during traditional fabric treatment. In this paper, cutinase-catalyzed green hydrolysis of WPET was achieved. The main purpose of this paper is to study the difference between two PEG-free WPETs with different -SO3- content in the cutinase-catalyzed hydrolysis process. Differences in solution properties, enzyme activity, SSIPA, and TPA release as well as functional groups and thermal properties of the hydrolysis products of the systems during cutinase hydrolysis were analyzed in detail for 14.1 % S-0 % PEG and 9.5 % S-0 % PEG. Results show that the structure of WPET has an influential effect on its hydrolysis rate in cutinase-catalyzed systems, which mainly originates from the -SO3- content. SSIPA released by WPET during hydrolysis has an inhibitory effect on cutinase enzyme activity. The hydrolysis process of WPET with higher -SO3- content releases more SSIPA, which in turn inhibits the enzyme activity of cutinase in the reaction system, ultimately leading to a slower hydrolysis rate of WPET. These findings provide new ideas for cutinase to target WPETs with different structures and are important for the green development of the textile industry.

Effect of PEG‐400 on dyeing properties of cotton fabric in non‐aqueous medium system

AbstractTraditional reactive dyeing has the technical problems of high water consumption and high pollution. In this article, the effect of polyethylene glycol 400 (PEG‐400) on the dyeing properties of cotton fabrics with high concentration reactive dyes in non‐aqueous medium was studied. Initially, the effects of PEG‐400 on colour strength (K/S) and colour fastness of dyed fabric with reactive dye are systematically discussed. In addition, the effect of PEG‐400 on the hydrolysis properties of reactive dyes was analysed by high‐performance liquid chromatography (HPLC). At the same time, the molecular interaction force between the dye and PEG‐400 was analysed by ultraviolet‐visible (UV‐visible) spectroscopy. The results showed that when PEG‐400 concentration was 3.0 g/L, K/S value of dyed fabric (5.0%, on weight of fabric, owf) was increased by 33.5%, and is much higher than dyed fabrics without PEG‐400. HPLC and UV‐visible analysis showed that the addition of PEG‐400 effectively reduced the hydrolysis rate of dye II containing two vinyl sulfone reactive groups, which was converted from C.I. Reactive Black 5 containing contains diethylsulfone sulphate group. Its hydrolysis rate was reduced by 20.9%. PEG‐400 also decreased the aggregation degree of high concentration reactive dye, and increased the solubility of reactive dye. This method has certain universality. Therefore, using PEG‐400 as an additive is an effective way to improve the dye uptake of reactive dyes in non‐aqueous medium dyeing system and realise cleaner production.

Microstructure of starch complexes coated with arabinoxylans and β-glucans: Effect on gelatinization and digestion properties

Research has shown that the presence of a complete cell wall structure in the grain endosperm reduces the digestibility of grains (starch) in the human body. In this study, composite structures of diverse contents of arabinoxylan (AX) and β-glucan (BG) in combination with wheat starch were constructed to simulate the grid structure formed by the combination of starch granules wrapped within cell walls in the starch endosperm. The gel properties following gelatinization and aging of the complexes were analyzed using rheology, differential scanning calorimetry, X-ray scanning, and infrared spectroscopy. Based on this, the differences in the in vitro digestion process of starch complexes were investigated, and the digestion mechanism was explored to offer a theoretical basis for the development of starch-based foods with a low glycemic index. The synergistic effect of AX and BG was found to promote the entanglement and aggregation of molecules, AX and BG inhibits the gelatinization of starch. AX/BG (10 % blend proportion) significantly inhibited starch digestion in the starch complexes, with the lowest hydrolysis rate (76.7 %), which may be because AX and BG wrap the starch particles effectively.

Discovery of (2R,4R)-4-((S)-2-Amino-3-methylbutanamido)-2-(4-boronobutyl)pyrrolidine-2-carboxylic Acid (AZD0011), an Actively Transported Prodrug of a Potent Arginase Inhibitor to Treat Cancer.

Arginase is a promising immuno-oncology target that can restore the innate immune response. However, it's highly polar active site often requires potent inhibitors to mimic amino acids, leading to poor passive permeability and low oral exposure. Using structure-based drug design, we discovered a novel proline-based arginase inhibitor (10) that was potent but had low oral bioavailability in rat. This issue was addressed by incorporating amino acids to target PepT1/2 active transport, followed by in vivo hydrolysis post absorption. The hydrolysis rate was highly tunable, and the valine prodrug (19) showed the best balance of stability and exposure of the potent payload. Dosing of 19 in mouse xenograft models significantly increased arginine in the tumor microenvironment, resulting in tumor growth inhibition as a monotherapy and in combination with an anti-PD-L1 antibody. Compound 19 (AZD0011) displays good pharmacokinetics and was selected as a clinical drug candidate for cancer.

Next-Gen Poly(ε-Caprolactone) Scaffolds: Non-Destructive In Vivo Monitoring and Accelerated Biodegradation.

Poly(ɛ-caprolactone) (PCL) is a biocompatible, biodegradable, and highly mechanically resilient FDA-approved material (for specific biomedical applications, e.g. as drug delivery devices, in sutures, or as an adhesion barrier), rendering it a promising candidate to serve bone tissue engineering. However, in vivo monitoring of PCL-based implants, as well as biodegradable implants in general, and their degradation profiles pose a significant challenge, hindering further development in the tissue engineering field and subsequent clinical adoption. To address this, photo-cross-linkable mechanically resilient PCL networks are developed and functionalized with a radiopaque monomer, 5-acrylamido-2,4,6-triiodoisophthalic acid (AATIPA), to enable non-destructive in vivo monitoring of PCL-based implants. The covalent incorporation of AATIPA into the crosslinked PCL networks does not significantly affect their crosslinking kinetics, mechanical properties, or thermal properties, but it increases their hydrolysis rate and radiopacity. Complex and porous 3D designs of radiopaque PCL networks can be effectively monitored in vivo. This work paves the way toward non-invasive monitoring of in vivo degradation profiles and early detection of potential implant malfunctions.

Synergistic effect of ozone treatment with α-amylase on the modification of microstructure and paste properties of japonica rice starch

The objective was to evaluate the synergistic effect of ozonization and α-amylase on modifying the microstructure and paste properties of starch, using 0.00042 g of ozone/100 g of buffer for various durations. Enzymatic susceptibility was increased, achieving maximum values of 12.73 % with an 11.42 % increase in crystallinity and an average particle size of 10.12 μm for 90 min treated japonica rice starch (JR90). The granules exhibited a polyhedral shape and, with increased intensity of combined treatments, formed clusters and lost their original geometry. Apparent viscosity, rheological, and textural parameters were reduced due to the more efficient action of α-amylase on ozonized starch, as confirmed by the low gelatinization enthalpy value (7.61 J/g). Ozone proved effective in opening starch chains, partially gelatinizing granules, homogenizing the enzymatic medium, and increasing the hydrolysis rate of α-amylase in japonica rice starch.

From Beer to Cheese: Characterization of Caseinolytic and Milk-Clotting Activities of Proteases Derived from Brewer's Spent Grain (BSG).

This study explores the extraction and characterization of proteolytic enzymes from brewer's spent grain (BSG) and their potential as sustainable coagulants in the dairy industry. BSG samples from various beer types (Blonde Ale, IPA, Kölsch, Honey, and Porter) were obtained from two artisanal breweries in Mar del Plata, Argentina. Optimization of caseinolytic activity (CA) and protein extraction was conducted using a Plackett-Burman design, followed by a Box-Behnken design. Optimal protein concentration was achieved at intermediate pH and high temperature, while CA peaked at pH 8.0. The specific caseinolytic activity (SCA) varied among the extracts, with BSG3 showing the highest activity (99.6 U mg-1) and BSG1 the lowest (60.4 U mg-1). Protease inhibitor assays suggested the presence of aspartic, serine, metallo, and cysteine proteases. BSG3 and BSG4 showed the highest hydrolysis rates for α-casein (70% and 78%). For κ-casein, BSG1, BSG2, and BSG3 demonstrated moderate activity (56.5%, 49%, and 55.8), while BSG4 and BSG5 exhibited the lowest activity. Additionally, the milk-clotting activity (MCA) of BSG extracts was comparable to plant-based coagulants like Cynara cardunculus and Ficus carica. These findings highlight the potential of BSG-derived proteases as alternative coagulants for cheese production, offering a sustainable link between the brewing and dairy industries.

Interactions Between Corn Starch and Ethyl Maltol Under Heat-Moisture Treatment and Its Application in Fried Chicken Nuggets.

This study delved into the interaction between corn starch and ethyl maltol during innovative repeated continuous heat-moisture treatment (RCHMT) and its impact on the quality of fried chicken nuggets. The results reveal that the complexation ratio of ethyl maltol is about 31.6%, and the complex creates dense microporous structures. Native starch and complex samples exhibited an A-type crystal structure, while the physical mixture sample showed superposition peaks of starch and ethyl maltol. Additionally, the peak of C-O-H def., CH2 of the complex sample was blue-shifted to the larger wave number, and the hydrogen bond structure was enhanced. Moreover, the complex exhibited a higher resistant starch content and lower hydrolysis rate and amylose content than the physical mixture sample. The starch-ethyl maltol complex has been demonstrated to be a non-inclusion compound. It has been shown to reduce oil absorption and enhance the crispness of fried chicken nuggets, matching that of commercial products. This finding provides a direction for the development of innovative coating powders.

Yeast β‒fructosidase-catalyzed synthesis of tyrosol fructoside in mono- and biphasic organic–aqueous systems

Abstract Tyrosol and its glycosides offer cell protection from oxidative stress and various health benefits. In this study, Saccharomyces cerevisiae β‒fructosidase potential for synthesizing tyrosol fructoside in mono- and biphasic solvent systems was assessed using 1.5 M sucrose, 10 g/L or 25 g/L tyrosol, at pH 6 and 40°C. Monophasic organic–water systems were designed with nine organic solvents (log P ranging from -1.35 to 0.5). Initial rates of sucrose hydrolysis, tyrosol and sucrose transfructosylation and tyrosol fructoside yields were evaluated. The hydrolytic activity increased with the addition of 5–15% organic solvents, revealing no clear correlation between the log P and sucrose hydrolysis rate. Simultaneously, transfructosylation reaction rates decreased, despite lower water activity. Notably, the effect of log P on tyrosol transfructosylation was observed in 70:30 (v/v) aqueous/organic biphasic systems. Differences in tyrosol fructoside synthesis in six biphasic reaction media were attributed to the unequal solubility of tyrosol in the organic phase, impacting its availability for the enzyme. Product yields were either similar to or lower than those in aqueous media, with no tyrosol fructoside partitioning into the organic phase, thus no effect of product stripping could be observed. Consequently, transfructosylation of tyrosol proves most effective in organic solvent-free media.

Impact of Flooding on Lands with Emerging Contaminants on the Quality of Receiving Water Bodies

Among crucial factors that control flooding events are extreme climate, urban growth, and mismanagement. Islands in the Nile River have experienced successive cycles of flooding and drying due to recent rainy years in the Nile Basin countries. This paper focuses on the impact of floods on the amounts of ammonium-N released, total concentrations of heavy metals, enzyme activities, and microbial biomass (C and N) in flood soils of 11 Nile River islands. Field moist soils and their airdried counterparts were collected from the outskirts of the island and incubated for 15 days at 30 °C to be analyzed. Results reflected that the amounts of released NH4-N were higher in airdried than in wet soils. The average hydrolysis rates of the studied six enzyme substrates were correlated significantly with organic C, organic N, microbial bio-mass C and N in the field moist soils. The highest correlation coefficients (r) were with rates of hydrolysis of substrates of asparaginase, amidase, and β-glycosaminidase. A significant intercorrelation between rates of hydrolysis of the six studied enzyme substrates and r values which range from 0.81 to 0.98. At 10 mmol kg−1 soil, Co+2, Cd+2, Pb+2, Cu+2, Cr+3, and Ni+2 inhibited the release of NH4-N. Arginine hydrolysis was inhibited by the six heavy metals at 5 mmol kg−1 soil; the most effective inhibitors were Pb+2, Ni+2, and Cr+3. Due to the complex soil enzymatic and organic nitrogen hydrolase activities that occur during floods, fertile fields on the Nile River islands should not be fertilized before the summer flood seasons, which will prevent contamination of the Nile River water and loss of its island soil fertility.

Fusion of Hydrophobic Anchor Peptides Promotes the Hydrolytic Activity of PETase but not the Extent of PET Depolymerization

AbstractEnzymatic recycling of polyethylene terephthalate (PET) has attracted significant attention in recent years. While the fusion of anchor peptides to PET hydrolases is believed to enhance PET hydrolytic activity, a quantitative analysis is yet lacking. Here, we construct four fusion enzymes by fusing anchor peptides (including hydrophobic LCI, LCIM1 and TA2, and hydrophilic EK4) to the C terminus of HotPETase, one of the most active PET hydrolases for high‐crystallinity PET (HC‐PET). Single‐molecule force spectroscopy (SMFS) demonstrates that hydrophobic anchor peptides promote adhesive interactions between the fusion enzymes and the PET surface. This is also validated by the adsorption kinetics and isotherms, and the saturated adsorption capacity remains unaltered compared to HotPETase. At low substrate loadings, the apparent hydrolytic activity of these fusion enzymes is positively related to the hydrophobicity of the anchor peptides. Among them, HotPETase‐LCI stands out as the most effective enzyme for HC‐PET degradation, demonstrating a 1.5‐fold increase in hydrolytic activity. At high substrate loadings, the advantages of fusion with anchor peptides diminish. We conclude that fusion enzymes only facilitate the hydrolytic rates of reactions for HC‐PET but have little effect on the final conversion extent.

Influence of the Conditions of Alkaline Pretreatment of Vegetable Raw Material before Hydrolysis in a Rotor Pulsation Device on the Cellulose Conversion

Introduction. The growing global energy demand, limited fossil fuel resources, and increasing greenhouse gasemissions highlight the need for expanded use of renewable energy sources, particularly biomass.Problem Statement. A significant drawback in the production of bioethanol is its high cost, primarily due tothe presence of hemicellulose and lignin.Purpose. This study aims to determine the effect of treating an alkaline suspension of wheat straw in a rotary pulsation device on the degree of cellulose conversion during enzymatic hydrolysis, a critical step in the productionof fuel ethanol.Materials and Methods. The raw material is pre-crushed wheat straw. The experiments have been conductedwith the use of a rotary pulsation device in an experimental plant.Results. It has been found that the combined physical effects of the discrete pulse energy input method andreduced straw particle size significantly increase the lignin extraction, from 40.0% at an average particle size of2—1 mm to 62.0% at an average particle size of 0.4—0.1 mm. Additionally, the treatment of an alkaline suspension of wheat straw at 90 оC for one hour, at alkali concentrations ranging from 1 to 4% (wt./wt.), results in anincrease in the cellulose conversion during enzymatic hydrolysis from 38% to 65.8%. Additionally, a corresponding decrease in the lignin content, from 17.1% to 3.16% has been reported.Conclusions. Increasing alkali concentration during the alkaline pretreatment of wheat straw using the discrete-pulse energy input method in a rotary pulsation device enhances the rate of cellulose conversion duringsubsequent enzymatic hydrolysis.

Catalytic hydrogenation of CO2 by composite nickel species supported on KCl-doped graphitic carbon nitride under normal temperature and atmospheric pressure

CO2 can be used as a cheap C1 feedstock to produce value-added chemical products and fuels. In this work, it was proposed for the first time that potassium chloride-doped graphitic carbon nitride-supported composite nickel species (CNS/KCl-g-C3N4) was prepared to catalyze CO2 hydrogenation with potassium borohydride (KBH4) as a hydrogen donor to produce formate at normal temperature and atmospheric pressure. The catalytic activity of CNS/KCl-g-C3N4 reached the best when the nickel loading was optimized as 15 wt%. By using CNS/KCl-g-C3N4, the hydrogen production volume and rate of KBH4 hydrolysis were reduced by 49 % and 32 %, respectively, while the formate concentration was increased by 43 %. It was speculated that the catalytic hydrogenation of CO2 was enhanced due to the chemical adsorption of CO2 on the catalyst surface by interacting with the K-N bonds and -NH2 groups, and the formation of BH4-n(OH)n- (n=0–3) with higher activity from BH4- on the Ni0 sites.

Differences in protein hydrolysis on quality of fermented milk: The changes in structure and processing characteristics

The functions and sensory properties of fermented dairy products are influenced by numerous factors, including the type of milk, the strain's degree of protein hydrolysis, and the additives of fermented milk. Currently, a large number of studies have demonstrated that the proteolytic ability of probiotics significantly affects the gel properties of fermented milk. However, few studies have examined the effects of probiotics with varying proteolytic abilities on the functions and sensory properties of fermented dairy products from the protein conformation perspective. Therefore, this study screened Lactobacillus helveticus KLDS1.8701 with a high protein hydrolysis rate and Lactobacillus helveticus KLDS1.0633 with a low hydrolysis rate to investigate the effects of varying degrees of protein hydrolysis on the gel properties, sensory properties and protein conformation of fermented milk. The results showed that the fermented milk containing Lactobacillus helveticus KLDS1.0633 with a low protein hydrolysis degree had high WHC, hardness and viscosity, a dense and uniform structure, abundant pores, and good texture and sensory properties. In addition, compared with fermented milk with high protein hydrolysis, low protein hydrolysis also increased the content of disulfide bonds, low surface hydrophobicity, and more hydrophilic structures such as a-helix. This study explains the impact of differences in protein hydrolysis degree among strains on fermented milk from a new perspective of protein secondary and tertiary conformations, which provides new insights for selecting suitable strains to produce fermented milk with better taste and nutrition.

N-Aldehyde-Modified Phosphatidylethanolamines generated by lipid peroxidation are robust substrates of N-Acyl Phosphatidylethanolamine Phospholipase D.

N-acyl phosphatidylethanolamine-hydrolyzing phospholipase D (NAPE-PLD) hydrolyzes phosphatidylethanolamines (PE) where the headgroup nitrogen has been enzymatically modified with acyl chains of four carbons or longer (N-acyl-PEs or NAPEs). The nitrogen headgroup of PE can also be non-enzymatically modified by reactive lipid aldehydes, thus forming N-aldehyde modified-PEs (NALPEs). Some NALPEs such as N-carboxyacyl-PEs are linked to PE via amide bonds similar to NAPEs, but others are linked by imine, pyrrole, or lactam moieties. Whether NAPE-PLD can hydrolyze NALPEs was unknown. We therefore characterized the major NALPE species formed during lipid peroxidation of arachidonic acid and linoleic acid and generated various NALPEs for characterization of their sensitivity to NAPE-PLD hydrolysis by reacting synthesized aldehydes with PE. We found that NAPE-PLD could act on NALPEs of various lengths and linkage types including those derived from PE modified by malondialdehyde (N-MDA-PE), butane dialdehyde (N-BDA-PE), 4-hydroxynonenal (N-HNE-PE), 4-oxo-nonenal (N-ONE-PE), 9-keto-12-oxo-dodecenoic acid (N-KODA-PE), and 15-E2-isolevuglandin (N-IsoLG-PE). To assess the relative preference of NAPE-PLD for various NALPEs versus its canonical NAPE substrates, we generated a substrate mixture containing roughly equimolar concentrations of the seven NALPEs as well as two NAPEs (N-palmitoyl-PE and N-linoleoyl-PE) and measured their rate of hydrolysis. Several NALPE species, including the N-HNE-PE pyrrole species, were hydrolyzed at a similar rate as N-linoleoyl-PE and many of the other NALPEs showed intermediate rates of hydrolysis. These results significantly expand the substrate repertoire of NAPE-PLD and suggest that it may play an important role in clearing products of lipid peroxidation in addition to its established role in the biosynthesis of N-acyl-ethanolamines.

EB3-informed dynamics of the microtubule stabilizing cap during stalled growth

Microtubule stability is known to be governed by a stabilizing GTP/GDP-Pi cap, but the exact relation between growth velocity, GTP hydrolysis and catastrophes remains unclear. We investigate the dynamics of the stabilizing cap through in vitro reconstitution of microtubule dynamics in contact with micro-fabricated barriers, using the plus-end binding protein GFP-EB3 as a marker for the nucleotide state of the tip. The interaction of growing microtubules with steric objects is known to slow down microtubule growth and accelerate catastrophes. We show that the lifetime distributions of stalled microtubules, as well as the corresponding lifetime distributions of freely growing microtubules, can be fully described with a simple phenomenological 1D model based on noisy microtubule growth and a single EB3-dependent hydrolysis rate. This same model is furthermore capable of explaining both the previously reported mild catastrophe dependence on microtubule growth rates and the catastrophe statistics during tubulin washout experiments.

Physicochemical and digestive characteristics of high internal phase emulsion based on pumpkin seed oil bodies prepared at different pHs

In this study, we prepared high internal phase emulsions based on pumpkin seed oil body (HIPEs-PSOB) with 83% oil phase at pH 6.5, pH 9.0 and pH 11.0. The pH 11.0-HIPE-PSOB contained almost oleosin of 14.5 kDa, while other HIPEs-PSOB had oleosin and extrinsic proteins. With the increase of extraction pHs, the content of bound water, immobilized water, particle size, viscosity, viscoelastisity and shear stress decreased, and the free water and isoelectric point of HIPEs-PSOB increased. During stomach digestion, the hydrolysis rate of extrinsic proteins of all HIPEs-PSOB was higher than that of oleosin, and the proteins were hydrolyzed to produce resistant peptides. CLSM showed that all HIPEs-PSOB coalesced during gastric and the droplets were significantly reduced and dispersed in the intestine. The total FFA release decreased with the increase in extraction pH. This study provided a theoretical basis for the investigation and application of OBs and HIPEs.

Allosteric modulation of the Lon protease via ssDNA binding and local charge changes.

The ATPase Associated with diverse cellular Activities (AAA+) family of proteases play crucial roles in cellular proteolysis and stress responses. Like other AAA+ proteases, the Lon protease is known to be allosterically regulated by nucleotide and substrate binding. Although it was originally classified as a DNA binding protein, the impact of DNA binding on Lon activity is unclear. In this study, we characterize the regulation of Lon by single-stranded DNA (ssDNA) binding and serendipitously identify general activation strategies for Lon. Upon binding to ssDNA, Lon's ATP hydrolysis rate increases due to improved nucleotide binding, leading to enhanced degradation of protein substrates, including physiologically important targets. We demonstrate that mutations in basic residues that are crucial for Lon's DNA binding not only reduce ssDNA binding but result in charge-specific consequences on Lon activity. Introducing negative charge at these sites induces activation akin to that induced by ssDNA binding, whereas neutralizing the charge reduces Lon's activity. Based on single molecule measurements, we find this change in activity correlated with changes in Lon oligomerization. Our study provides insights into the complex regulation of the Lon protease driven by electrostatic contributions from either DNA binding or mutations.

Influence of replacing La or Mg with Ca on the hydrolysis of LaMg12H27 for hydrogen production

In this work, LaMg12H27, La0.2Mg12Ca0.8H26.2, La0.2Mg10Ca2.8H26.2, LaMg10Ca2H27, and LaMg6Ca6H27 samples were prepared by varying the milling time and replacing metal La or Mg with different metal Ca contents, and the hydrogen production properties of the samples were investigated and compared. The phase composition and morphology of the composites were analyzed using XRD and SEM. The results showed that the hydrogen conversion rate of the LaMg10Ca2H27 ball-milled for 10 h was 97%, while the hydrogen yield of the LaMg6Ca6H27 sample ball-milled for 10 h was 890 mL/g within 1 min. The analysis of the hydrolysis properties and mechanism of the samples revealed that the increase in hydrogen production was mainly attributed to the decrease in particle size and the increase in surface cracks of the samples after ball milling. It was also observed that an increase in the metal Ca content increased the initial hydrolysis rate of the composites, while a moderate substitution of the metal La increased the hydrogen production rate.

Efficient Biosynthesis of Theanderose, a Potent Prebiotic, Using Amylosucrase from Deinococcus deserti.

The study aimed to develop an efficient bioprocess for the discovery and synthesis of theanderose by using amylosucrase from Deinococcus deserti (DdAS). An unknown trisaccharide produced by DdAS was detected by high-performance anion-exchange chromatography-pulsed amperometric detection and high-performance liquid chromatography-evaporative light scattering detection, purified using medium-pressure liquid chromatography, and identified as theanderose (α-d-glucopyranosyl-(1→6)-α-d-glucopyranosyl-(1→2)-β-d-fructofuranoside) through nuclear magnetic resonance and mass spectrometry. DdAS synthesized theanderose with a 25.4% yield (174.1 g/L) using 2.0 M sucrose at 40 °C for 96 h. In an in vitro digestion model, theanderose showed a 6.5% hydrolysis rate over 16 h. Prebiotic efficacy tests confirmed that theanderose significantly enhanced the proliferation of selected Bifidobacterium strains in the culturing medium with theanderose as the main carbon source. Subsequently, fecal fermentation was performed by adding theanderose to the feces of 20 individuals of varying ages to assess its effect on the gut microbiota. Theanderose increased the relative abundance of Bifidobacteriaceae and Prevotellaceae while decreasing the population ratio of Lachnospiraceae and Ruminococcaceae. Conclusively, theanderose displayed excellent prebiotic potential when judged by low digestibility and selective growth of beneficial microbes over harmful microbes.