“…Kinetic parameters for each of the three substrates were also determined according to the Michaelis-Menten equation (Table 2), with V max values similar in magnitude and trend to the specific activities for each of the three main substrates (poly-ManA, poly-GlcA, and HA); V max for poly-GlcA is ϳ10-fold greater versus poly-ManA or HA. The K m values for the three substrates also follow a similar trend to V max and specific activity, with a 5-fold larger K m for HA versus poly-GlcA (30,47). Thus, based on the highest specific activity for poly-GlcA (Fig.…”
Section: Heterologous Expression and One-step Purification Ofsupporting
confidence: 55%
“…The highest overall specific activity (848.3 Ϯ 6.3 units/mg at pH 7) was for poly-GlcA (Fig. 5A), which is among the highest reported for poly-GlcAspecific lyases (30,44,45). For poly-ManA the specific activity (68.5 Ϯ 2.9 units/mg at pH 9) was also significant (Fig.…”
Section: Heterologous Expression and One-step Purification Ofmentioning
Background: Polysaccharide lyases degrade anionic polysaccharides and are important in bacterial biofilm formation and host-pathogen interactions. Results: Putative alginate lyase (Smlt1473) from Stenotrophomonas maltophillia displays activity toward multiple polysaccharides in a pH-regulated manner. Conclusion: Smlt1473 is a unique polysaccharide lyase with pH-dependent activity toward mammalian, plant, and microbial substrates. Significance: Characterization of Smlt1473 allows for an understanding of possible roles in biofilm formation and pathogenesis.
“…Kinetic parameters for each of the three substrates were also determined according to the Michaelis-Menten equation (Table 2), with V max values similar in magnitude and trend to the specific activities for each of the three main substrates (poly-ManA, poly-GlcA, and HA); V max for poly-GlcA is ϳ10-fold greater versus poly-ManA or HA. The K m values for the three substrates also follow a similar trend to V max and specific activity, with a 5-fold larger K m for HA versus poly-GlcA (30,47). Thus, based on the highest specific activity for poly-GlcA (Fig.…”
Section: Heterologous Expression and One-step Purification Ofsupporting
confidence: 55%
“…The highest overall specific activity (848.3 Ϯ 6.3 units/mg at pH 7) was for poly-GlcA (Fig. 5A), which is among the highest reported for poly-GlcAspecific lyases (30,44,45). For poly-ManA the specific activity (68.5 Ϯ 2.9 units/mg at pH 9) was also significant (Fig.…”
Section: Heterologous Expression and One-step Purification Ofmentioning
Background: Polysaccharide lyases degrade anionic polysaccharides and are important in bacterial biofilm formation and host-pathogen interactions. Results: Putative alginate lyase (Smlt1473) from Stenotrophomonas maltophillia displays activity toward multiple polysaccharides in a pH-regulated manner. Conclusion: Smlt1473 is a unique polysaccharide lyase with pH-dependent activity toward mammalian, plant, and microbial substrates. Significance: Characterization of Smlt1473 allows for an understanding of possible roles in biofilm formation and pathogenesis.
“…We isolated a bacterial strain, Brevundimonas sp. strain SH203, that is able to degrade cellouronate (23,28), and we purified two types of cellouronate lyase, cellouronate lyases I and II, which catalyze endo-and exodepolymerization of cellouronate, respectively. A combination of these two enzymes, moreover, resulted in synergistic degradation of cellouronate to its monomers (27,28).…”
mentioning
confidence: 99%
“…SEC was performed with a high-performance liquid chromatography system consisting of a Shodex Ohpak SB-802.5 HQ column (0.8 by 30 cm; Showa Denko Co., Tokyo, Japan) as described previously (27,39), and detection was carried out by determining both the refractive index (RID-10A; Shimadzu) and the UV absorption at 235 nm (SPD-M20A photodiode array; Shimadzu). 13 C NMR spectra of the products were collected with a Bruker AC-300 spectrometer as described previously (28).…”
The filamentous fungus Trichoderma reesei produces glucuronan lyase (TrGL) when it is grown on -(134)-polyglucuronate (cellouronate) as a sole carbon source. The cDNA encoding TrGL was cloned, and the recombinant enzyme was heterologously expressed in Pichia pastoris. The cDNA of TrGL includes a 777-bp open reading frame encoding a 20-amino-acid signal peptide and the 238-amino-acid mature protein. The amino acid sequence showed no similarity to the amino acid sequences of previously described functional proteins, indicating that the enzyme should be classified in a novel polysaccharide lyase (PL) family. Recombinant TrGL catalyzed depolymerization of cellouronate endolytically by -elimination and was highly specific for cellouronate. The enzyme was most active at pH 6.5 and 50°C, and its activity and thermostability increased in the presence of Ca 2؉ , suggesting that its calcium dependence is similar to that of other PLs, such as pectate lyases.
“…This characteristic opens the way to applications in gasbarrier biomaterial and constitutes environmentally friendly biodegradable packaging films. Studies have led to the biodegradation of these synthetic polyuronides using new specific polysaccharide cleavage enzymes as polysaccharide lyases or hydrolases in order to consider large productions of potential bioactive anionic oligosaccharides [55,56]. The use of these enzymatic activities could also generate new oligosaccharides with biological activities as described on lots of organisms such as bacteria, fungi, plant, algae and mammalian [57,58].…”
Section: Tempo/nabr/naocl System Mediated Oxida-tion Of Polysaccharidesmentioning
Abstract:Polyuronides are an acidic class of polysaccharides with interesting rheological and biological properties. However, except pectin and alginate, the structural variability of this class of polysaccharides is poor and low described in literature. In this context, a new generation of polyuronides has been isolated from two sources in the middle of the 90's. Firstly, a bacterial -(1,4) polyglucuronic acid called glucuronan was identified as the sole exopolysaccharide produced by a bacteria belonging to the Rhizobiaceae family. Secondly, the development of the TEMPO chemistry led to the production at large scale of oxidized cellulose called cellouronate. Both new polyuronides were largely patented and found applications in several industrial areas. Moreover, the biodegradation study of these polysaccharides has led to the identification of a new family of polysaccharide lyases very specific for these substrates. This review focuses on the actual knowledge of this class of acidic polysaccharides and on the enzymes acting about them.
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