Cloning, expression, and characterization of the β-glucosidase hydrolyzing secoisolariciresinol diglucoside to secoisolariciresinol from Bacteroides uniformis ZL1

Tao, Yilun; Yang, Donghui; Zhang, Y.-T.; Zhang, Y.; Wang, Z.-Q.; Wang, Y.-S.; Cai, Shao‐Qing; Liu, S.-L.

doi:10.1007/s00253-013-5111-7

Cited by 24 publications

(12 citation statements)

References 55 publications

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“…have been found to activate a variety of plant metabolites, including the lignin secoisolariciresinol diglucoside (SDG) (Clavel et al, 2006) and the flavonoid glycoside rutin (Bokkenheuser et al, 1987). The β-glucosidase involved in SDG hydrolysis in Bacteroides uniformis ZL1 was also found to exhibit activity on other plant glycosides, including astragalin, rutin, and isoquercetin (Tao et al, 2014). Similarly, an α-L-rhamnosidase from Bacteroides JY-6 was isolated for its activity on different rhamnoglucosides, including rutin, poncirin, and hesperidin (Jang and Kim, 1996).…”

Section: Discussionmentioning

confidence: 99%

A metabolic pathway for glucosinolate activation by the human gut symbiontBacteroides thetaiotaomicron

Liou

Sirk

Diaz

et al. 2019

Preprint

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Diet is the largest source of plant-derived metabolites that influence human health. The gut microbiota can metabolize these molecules, altering their biological function. However, little is known about the gut bacterial pathways that process plant-derived molecules. Glucosinolates are well-known metabolites in brassica vegetables and metabolic precursors to cancer-preventive isothiocyanates. Here, we identify a genetic and biochemical basis for isothiocyanate formation by Bacteroides thetaiotaomicron, a prominent gut commensal species. Using a genome-wide transposon insertion screen, we identified an operon required for glucosinolate metabolism in B. thetaiotaomicron. Expression of BT2159-BT2156 in a non-metabolizing relative, Bacteroides fragilis, resulted in gain of glucosinolate metabolism. We show that isothiocyanate formation requires the action of BT2158 and either BT2156 or BT2157 in vitro. Monocolonization of mice with mutant BtΔ2157 showed reduced isothiocyanate production in the gastrointestinal tract. These data provide insight into the mechanisms by which a common gut bacterium processes an important dietary nutrient.with the induction of Nrf2, ITC have also been shown to interact with sensory and inflammatory pathways. Allyl ITC, derived from the GS sinigrin, is an agonist of the transient receptor potential ion channel TRPA1, which is involved in signaling inflammatory pain (Bellono et al., 2017), while sulforaphane has been shown to covalently modify lipopolysaccharide-activated Toll-like receptor 4 (TLR4), resulting in the reduced secretion of pro-inflammatory cytokines in human peripheral blood mononuclear cells and monocytes (Folkard et al., 2014).Plant and gut bacterial metabolism have been identified as two different routes for the conversion of GS into ITC products required for biological activity. Like many phytonutrients relevant to human health, GS in crucifers are involved in chemical defense in planta (Halkier and Gershenzon, 2006). These metabolites are stored in the plant tissue in an inert, pro-drug-like state and are converted into an active state by myrosinase enzymes, thio-specific glucoside hydrolases found in cruciferous vegetables (Figure 1A). GS are sequestered in vacuoles under normal conditions, while myrosinases are expressed in distinct, specialized myrosin cells (Kissen et al., 2009). Upon disruption of the plant tissue, myrosinases mediate the hydrolysis of GS into a variety of possible products, including ITC, nitriles, and epithionitriles, determined by environmental pH and the presence of specifier proteins.

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Section: Discussionmentioning

confidence: 99%

A metabolic pathway for glucosinolate activation by the human gut symbiontBacteroides thetaiotaomicron

Liou

Sirk

Diaz

et al. 2019

Preprint

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“…Enzymatic cell breakage can also be adapted on a large scale, but this method was hindered by the cost of enzymes used. Meanwhile, most β-glucosidases produced in E. coli exist as intracellular proteins that require cell breakage to obtain the target protein [ 10 , 11 , 13 , 35 , 36 ]. Therefore, autolysis using proteins such as SRRz has attracted much attention during recent years.…”

Section: Discussionmentioning

confidence: 99%

“…IPTG is expensive, toxic, and difficult to remove during downstream operations [ 9 ]. Another obstacle on the production of β -glucosidases using E. coli is that they are often expressed as intracellular proteins [ 10 – 13 ]. Cell wall breakage is required in order to obtain the target protein.…”

Section: Introductionmentioning

confidence: 99%

Light induced expression of β-glucosidase in Escherichia coli with autolysis of cell

Fei

Zhang

et al. 2017

BMC Biotechnol

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Backgroundβ-Glucosidase has attracted substantial attention in the scientific community because of its pivotal role in cellulose degradation, glycoside transformation and many other industrial processes. However, the tedious and costly expression and purification procedures have severely thwarted the industrial applications of β-glucosidase. Thus development of new strategies to express β-glucosidases with cost-effective and simple procedure to meet the increasing demands on enzymes for biocatalysis is of paramount importance.ResultsLight activated cassette YF1/FixJ and the SRRz lysis system were successfully constructed to produce Bgl1A(A24S/F297Y), a mutant β-glucosidase tolerant to both glucose and ethanol. By optimizing the parameters for light induction, Bgl1A(A24S/F297Y) activity reached 33.22 ± 2.0 U/mL and 249.92 ± 12.25 U/mL in 250-mL flask and 3-L fermentation tank, respectively, comparable to the controls of 34.02 ± 1.96 U/mL and 322.21 ± 10.16 U/mL under similar culture conditions with IPTG induction. To further simplify the production of our target protein, the SRRz lysis gene cassette from bacteriophage Lambda was introduced to trigger cell autolysis. As high as 84.53 ± 6.79% and 77.21 ± 4.79% of the total β-glucosidase were released into the lysate after cell autolysis in 250 mL flasks and 3-L scale fermentation with lactose as inducer of SRRz. In order to reduce the cost of protein purification, a cellulose-binding module (CBM) from Clostridium thermocellum was fused into the C-terminal of Bgl1A(A24S/F297Y) and cellulose was used as an economic material to adsorb the fusion enzyme from the lysate. The yield of the fusion protein could reach 92.20 ± 2.27% after one-hour adsorption at 25 °C.ConclusionsWe have developed an efficient and inexpensive way to produce β-glucosidase for potential industrial applications by using the combination of light induction, cell autolysis, and CBM purification strategy.Electronic supplementary materialThe online version of this article (10.1186/s12896-017-0402-1) contains supplementary material, which is available to authorized users.

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“…Interestingly, B. uniformis expresses a gene that encodes β-glucosidase that releases glucose from SDG via hydrolysis 18,19 . We therefore feed mice with flaxseed lignans, which are high in SDG, and examined the proliferation of B. uniformis in the gut.…”

Section: Administration Of α-Cyclodextrin Increased B Uniformis Cellmentioning

confidence: 99%

Bacteroides uniformis enhances endurance exercise performance through gluconeogenesis

Morita¹,

Kano²,

Ishii

et al. 2020

Preprint

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Athletes require high levels of energy to exercise under extreme conditions. Gut microbiota supplies energy to the host; however, the mechanism how gut microbiota contribute in the athlete is unclear. In this study, we determined that gut microbiota of Japanese long-distance runners differed from that of non-athletes, and the Bacteroides uniformis cell number in the feces correlated with 3,000-m race time. Mice administrated with B. uniformis extended the swimming time to exhaustion. Furthermore, acetate and propionate concentrations in the cecum increased in B. uniformis-administered mice subjected to weekly exercise.Expression levels of carnitine palmitoyl transferase 1a and phosphoenolpyruvate carboxykinase genes were elevated in the liver, suggesting that acetate and propionate produced by B. uniformis improve endurance exercise performance, at least in part, through enhancing gluconeogenesis. In addition, α-cyclodextrin administration increased B. uniformis and improved the performance in humans and mice, thus it is a candidate substance enhancing exercise performance through modification of gut microbiota. INTRODUCTIONHuman exercise performance comprises a combination of physical strength (e.g., muscle and explosive strength, endurance, and flexibility) and the actions performed in a given exercise (e.g., throwing, kicking, and hitting). Exercise performance can be improved through training; however, other factors, such as genetic background, may affect it. Recently, several reports have demonstrated that the gut microbiota contributes to health, energy homeostasis, and glucose and lipid metabolisms 1-4 . Hsu et al. reported that the swimming endurance time of germ-free mice was shorter than that of specific-pathogen-free mice and Bacteroides fragilis-monoassociated mice, thus identifying gut microbiota as one of the factors associated with exercise performance 5 . Conversely, other studies have shown that exercise changes the gut microbiome profile in mice 6,7 . In humans, Clarke et al. reported that the gut microbiota of rugby players had high α diversity and relative abundance of the genera Prevotella, Succinivibrio, Bacteroides, and Lactobacillus, as a difference to those of non-rugby players 8 . Another study on the gut microbiota of cyclists showed that a high abundance ofPrevotella was correlated with weekly exercise time 9 . However, none of these studies demonstrated a causal relationship between exercise performance and characteristic genera in the gut microbiota of athletes. Recently, Veillonella atypica, isolated from the gut microbiota of marathon runners, was shown to produce propionate from exercise-induced lactate in the gut, improving endurance exercise performance in mice 10 . Only that report has demonstrated a causal relationship between exercise performances and the gut microbiota in mice, but 3 detailed relationship remains unclear. In the present study, we investigated the role of Bacteroides uniformis, identified in the microbiome analysis of Japanese long-distance runners, in the en...

show abstract

Cloning, expression, and characterization of the β-glucosidase hydrolyzing secoisolariciresinol diglucoside to secoisolariciresinol from Bacteroides uniformis ZL1

Cited by 24 publications

References 55 publications

A metabolic pathway for glucosinolate activation by the human gut symbiontBacteroides thetaiotaomicron

A metabolic pathway for glucosinolate activation by the human gut symbiontBacteroides thetaiotaomicron

Light induced expression of β-glucosidase in Escherichia coli with autolysis of cell

Bacteroides uniformis enhances endurance exercise performance through gluconeogenesis

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