Enzymatic Degradation of Allergen Peptides from Bovine Casein by a Combination of Streptomyces Aminopeptidases

Wan, Kun; Uraji, Misugi; Tokai, Shota; Hatanaka, Tadashi

doi:10.1007/s12010-018-2839-7

Cited by 15 publications

(5 citation statements)

References 28 publications

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“…We have investigated Streptomyces APs for more than 10 years (Arima et al 2004(Arima et al , 2006(Arima et al , 2008Hatanaka et al 2007Hatanaka et al , 2011Uraji et al 2007) and recently evaluated the synergistic effects of three Streptomyces APs on casein hydrolysis to degrade VLPVPQK and FFVAPFPEVFGK, which are known as allergenic and bitter peptides derived from tryptic casein (Chen et al 2015;Weber et al 2006;Nishiwaki et al 2002). An APP from Streptomyces costaricanus TH-4 (Arima et al 2008) was found to release the N-terminal Val from the sequence VLPVPQK (Wan et al 2019). This result indicates that the APP from Streptomyces may exhibit a novel substrate specificity.…”

Section: Introductionmentioning

confidence: 93%

“…The procedure was similar to a previously described method (Wan et al 2019). A mixture of 20 mg trypsin (225 proteolytic units/mg), 2 g of casein, and 200 mL of 50 mM ammonium bicarbonate (NH 4 HCO 3 , pH 7.0) was incubated at 40 °C overnight on a rotary shaker at 750 rpm; the mixture was then incubated at 90 °C for 30 min without shaking to inactivate trypsin.…”

Section: Preparation Of Tryptic Caseinmentioning

confidence: 99%

“…The procedure was similar to a previously described method (Wan et al 2019). In brief, a mixture of 33 µL of Bioprase SP (100 proteolytic units/mg, 10 mg/mL) and 10 mL of 0.6% (w/v) aqueous dispersion of casein (40 mM monosodium phosphate [NaH 2 PO 4 ], pH 7.5) was prepared and incubated at 40 °C overnight and then at 80 °C for 30 min to inactivate Bioprase SP.…”

Section: Preparation Of Casein Hydrolysate and Analysis Of Free Aminomentioning

confidence: 99%

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Novel activity of Streptomyces aminopeptidase P

Wan

Uraji

Yang³

et al. 2020

Bioresour. Bioprocess.

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Streptomyces aminopeptidase P enzymes are proline-specific peptidases that belong to the peptidase M24 family. To evaluate the activity of a commercial Streptomyces aminopeptidase P, named 'XPO DUET' , we performed three experiments involving degradation of tryptic casein, production of free amino acids from casein hydrolysate, and hydrolysis of synthetic peptides. Using an ion-trap liquid chromatography-mass spectrometry (LC-MS) apparatus, we demonstrate that XPO DUET could degrade FFVAPFPEVFGK, an allergic and bitter peptide, VAPFPEVFGK, and PEVFGK from tryptic casein. All amino acids, except Ala, Asp, Glu, and Tyr, were released in an XPO DUET activity-dependent manner during the hydrolysis of casein hydrolysate. LC-MS analysis also revealed the ability of XPO DUET to completely hydrolyze Phe-Phe-Phe into free Phe. Thus, we confirm that XPO DUET possesses broader specificity than its known activity toward Xaa-Pro peptides. Because XPO DUET is a food-grade peptidase, it is useful in the bioprocessing of protein hydrolysates through its combination with other food-grade peptidases.

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Section: Introductionmentioning

confidence: 93%

Section: Preparation Of Tryptic Caseinmentioning

confidence: 99%

Section: Preparation Of Casein Hydrolysate and Analysis Of Free Aminomentioning

confidence: 99%

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Novel activity of Streptomyces aminopeptidase P

Wan

Uraji

Yang³

et al. 2020

Bioresour. Bioprocess.

Self Cite

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“…Recently, milk‐derived EV have been under intensive scrutiny as delivery vehicles (Rashidi et al., 2022 ) and show unique advantages as oral delivery tools for gastrointestinal diseases due to the biocompatibility, resistance to gastrointestinal microenvironment and accessibility (Zhong et al., 2021 ). However, unlike EV derived from cell culture, casein is one of main contaminants during the extraction process of EV from milk (Wan et al., 2019 ) and UC patients commonly suffer from milk intolerance (Judaki et al., 2014 ), removal of casein is critical for purification of EV designed for oral administration. Although different approaches have been explored including dissociation of casein with the chelator ethylenediamine tetraacetic acid (EDTA), precipitation via acidification (Rahman et al., 2019 ; Vaswani et al, 2019 ; Welsh et al, 2024 ) or ultracentrifugation (Munagala et al., 2016 ), there is no consensus on the production methodology and there is room for further improving the purity of EV.…”

Section: Introductionmentioning

confidence: 99%

Milk‐derived extracellular vesicles functionalized with anti‐tumour necrosis factor‐α nanobody and anti‐microbial peptide alleviate ulcerative colitis in mice

Jing,

Zhang,

et al. 2024

J of Extracellular Vesicle

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Ulcerative colitis (UC) manifests clinically with chronic intestinal inflammation and microflora dysbiosis. Although biologics can effectively control inflammation, efficient delivery to the colon and colon epithelial cells remains challenging. Milk‐derived extracellular vesicles (EV) show promise as an oral delivery tool, however, the ability to load biologics into EV presents challenges to therapeutic applications. Here, we demonstrate that fusing cell‐penetrating peptide (TAT) to green fluorescent protein (GFP) enabled biologics loading into EV and protected against degradation in the gastrointestinal environment in vitro and in vivo after oral delivery. Oral administration of EV loaded with anti‐tumour necrosis factor‐α (TNF‐α) nanobody (VHHm3F) (EVVHH) via TAT significantly reduced tissue TNF‐α levels and alleviated pathologies in mice with acute UC, compared to VHH alone. In mice with chronic UC, simultaneously introducing VHH and an antimicrobial peptide LL37 into EV (EVLV), then administering orally improved intestinal barrier, inflammation and microbiota balance, resulted in relief of UC‐induced depression and anxiety. Collectively, we demonstrated that oral delivery of EVLV effectively alleviated UC in mice and TAT efficiently loaded biologics into EV to confer protection from degradation in the gastrointestinal tract. This therapeutic strategy is promising for UC and is a simple and generalizable approach towards drug‐loaded orally‐administrable EV treatment for other diseases.

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“…The vector harbors ori and rep for streptomycetes and thiostrepton-resistant genes (Hatanaka et al 2008). By combining this vector with the S. lividans 1326 strain, we succeeded in the extracellular production of several Streptomyces enzymes (Hatanaka et al 2008;Uraji et al 2018;Wan et al 2019). In this study, we attempted to identify an essential region of the scmp promoter to enhance its expression.…”

Section: Introductionmentioning

confidence: 99%

Improvement in Hyperexpression with a Combination of Truncated Scmp Promoter and Streptomyces Lividans

Uraji

L²,

Hatanaka³

2022

Preprint

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We have previously reported a powerful promoter from the Streptomyces cinnamoneus TH-2 strain named ‘scmp’ and created an expression vector of pTONA5a based on pIJ702 for expression using S. lividans (Hatanaka et al. (2008) Protein Expr Purif 62: 244-248). The full-length scmp promoter sequence consists of 424 bp upstream of a metalloendoprotease gene in the S. cinnamoneus TH-2 genome. The promoter works in the presence of inorganic phosphate and glucose. In this study, we present the essential region of the scmp promoter (promoter C), which lacks 358 bp of the 5′ region of the full-length promoter. Promoter C was very short and contained only 63 bp. Moreover, there was no restriction on the medium component by combining the promoter and the S. livians 1326 strain. Using promoter C and S. lividans, we succeeded in the extracellular production of the Streptomyces enzymes leucine aminopeptidase, ferulic acid esterase, and transglutaminase, which possessed signal peptides for secretion via the second pathway, at high levels.

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Enzymatic Degradation of Allergen Peptides from Bovine Casein by a Combination of Streptomyces Aminopeptidases

Cited by 15 publications

References 28 publications

Novel activity of Streptomyces aminopeptidase P

Novel activity of Streptomyces aminopeptidase P

Milk‐derived extracellular vesicles functionalized with anti‐tumour necrosis factor‐α nanobody and anti‐microbial peptide alleviate ulcerative colitis in mice

Improvement in Hyperexpression with a Combination of Truncated Scmp Promoter and Streptomyces Lividans

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