Identification and characterization of a PL35 GAGs lyase with 4-O-sulfated N-acetylgalactosamine (A-type)-rich structures producing property.

Glycosaminoglycan (GAG) lyases are important tools for investigating the structure of GAGs and preparing low-molecular-weight GAGs. The PL35 family, a recently established polysaccharide lyase family, should be further investigated. In this study, we discovered a new GAG lyase, CHa1, which belongs to the PL35 family. When expressed heterologously in Escherichia coli (BL21), CHa1 exhibited high expression levels and solubility. The optimal activity was observed in Tris-HCl buffer (pH 7.0) or sodium phosphate buffer (pH 8.0) at 30 °C. The specific activities towards HA, CSA, CSC, CSD, CSE, and HS were 3.81, 13.03, 36.47, 18.46, 6.46, and 0.50 U/mg protein, respectively. CHa1 digests substrate chains randomly that acting as an endolytic lyase and shows a significant preference for GlcA-containing structures, prefers larger oligosaccharides (≥UDP8) and can generate a series of oligosaccharides composed mainly of the A unit when digesting CSA. These oligosaccharides include ΔC-A, ΔC-A-A, ΔC-A-A-A, ΔC-A-A-A-A, and ΔC-A-A-A-A-A. The residues Tyr257 and His421 play crucial roles in the catalytic process, and Ser211, Asn212, Asn213, Trp214, Gln216, Lys360, Arg460 and Gln462 may participate in the binding process of CHa1. This study on CHa1 contributes to our understanding of the PL35 family and provides valuable tools for investigating the structure of GAGs.

Key processes in tumor metastasis and therapeutic strategies with nanocarriers: a review.

Cancer metastasis is the leading cause of cancer-related death. Metastasis occurs at all stages of tumor development, with unexplored changes occurring at the primary site and distant colonization sites. The growing understanding of the metastatic process of tumor cells has contributed to the emergence of better treatment options and strategies. This review summarizes a range of features related to tumor cell metastasis and nanobased drug delivery systems for inhibiting tumor metastasis. The mechanisms of tumor metastasis in the ideal order of metastatic progression were summarized. We focus on the prominent role of nanocarriers in the treatment of tumor metastasis, summarizing the latest applications of nanocarriers in combination with drugs to target important components and processes of tumor metastasis and providing ideas for more effective nanodrug delivery systems.

Assessment of soil quality in an arid and barren mountainous of Shandong province, China.

Forest soils are important components of forest ecosystems, and soil quality assessment as a decision-making tool to understand forest soil quality and maintain soil productivity is essential. Various methods of soil quality assessment have been developed, which have occasionally generated inconsistent assessment results between soil types. We assessed the soil quality of five communities (herb, shrub, Quercus acutissima, Pinus thunbergii, and Q. acutissima-P. thunbergii mixed plantation) using two common methods of dry and barren mountains in the Yimeng Mountain area, China. Sixteen soil physical, chemical and biological properties were analysed. The soil quality index was determined using the established minimum data set based on the selection results of principal component analysis and Pearson analysis. Silt, soil total phosphorus (P), soil total nitrogen (N), L-leucine aminopeptidase, acid phosphatase and vector length were identified as the most representative indicators for the minimum data set. Linear regression analysis showed that the minimum data set can adequately represent the total data set to quantify the impact of different communities on soil quality (P < 0.001). The results of linear and non-linear methods of soil quality assessment showed that the higher soil quality index was Pinus forest (0.59 and 0.54), and the soil quality index of mixed plantation (0.41 and 0.45) was lower, which was similar to the herb community (0.37 and 0.44). Soil quality was mostly affected by soil chemical properties and extracellular enzyme activities of different communities, and the different reasons for the low soil quality of mixed plantations were affected by soil organic carbon (C) and total C. Overall, we demonstrate that the soil quality index based on the minimum data set method could be a useful tool to indicate the soil quality of forest systems. Mixed plantations can improve soil quality by increasing soil C, which is crucial in ecosystem balance.

Identification and characterization of a novel heparinase PCHepII from marine bacterium Puteibacter caeruleilacunae.

Heparin (HP) and heparan sulfate (HS) are multifunctional polysaccharides widely used in clinical therapy. Heparinases (Hepases) are enzymes that specifically catalyse HP and HS degradation, and they are valuable tools for studying the structure and function of these polysaccharides and for preparing low molecular weight heparins. In this study, by searching the NCBI database, a novel enzyme named PCHepII was discovered in the genome of the marine bacterium Puteibacter caeruleilacuae. Heterologously expressed PCHepII in Escherichia coli (BL21) has high expression levels and good solubility, active in sodium phosphate buffer (pH 7.0) at 20°C. PCHepII exhibits an enzyme activity of 254 mU/mg towards HP and shows weak degradation capacity for HS. More importantly, PCHepII prefers to catalyse the high-sulfated regions of HP and HS rather than the low-sulfated regions. Although PCHepII functions primarily as an endolytic Hepase, it mainly generates disaccharide products during the degradation of HP substrates over time. Investigations reveal that PCHepII exhibits a preference for catalysing the degradation of small substrates, especially HP tetrasaccharides. The catalytic sites of PCHepII include the residues His199, Tyr254, and His403, which play crucial roles in the catalytic process. The study and characterization of PCHepII can potentially benefit research and applications involving HP/HS, making it a promising enzyme.

Simulation and Experimental Study on the Precision Molding of Irregular Vehicle Glass Components.

The high level of stress and dimension deviation induced by glass molding are the main causes of the low yield rate of large, irregular glass components on vehicles. To solve this issue, a numerical model of large glass component molding was established in this study, which aimed to analyze the dominant factors of molding quality and achieve a synergistic balance between quality characteristics and energy consumption. The results show that molding temperature is the dominant factor affecting the energy consumption and residual stress, and the molding pressure is the main factor affecting the dimension deviation. Furthermore, the NSGA-II optimization algorithm was used to optimize the maximum residual stress, dimension deviation, and energy consumption with the numerical results. The combination of a heating rate of 1.95 °C/s, holding time of 158 s, molding temperature of 570 °C, molding pressure of 34 MPa, and cooling rate of 1.15 °C/s was determined to be the optimized scheme. The predictive error of the numerical result, based on the optimized scheme, was experimentally verified to be less than 20%. It proved the accuracy of the model in this study. These results can provide guidance for the subsequent precision molding of large, irregular glass components.

Water quality improvement project for initial rainwater pollution and its performance evaluation.

Efficiently addressing initial rainwater pollution is crucial for mitigating urban water pollution. However, the performance evaluation of initial rainwater pollution control project is rarely introduced. In this study, the architecture of effective comprehensive engineering measures for improving the water quality of initial rainwater in Anhui Province, China, was described. Three water quality indicators, ammonia nitrogen (NH3-N), chemical oxygen demand (COD), and total phosphorus (TP), were selected to explore the severity of urban pollution caused by initial rainwater under various rainfall scenarios. A single-factor evaluation method was used to contrast and assess the benefits of the initial rainfall interception project in terms of water quality enhancement. Results showed that initial rainfall pollution was gentler under light rainfall conditions but more prominent under moderate and heavy conditions. The percentages of NH3-N, COD, and TP in Lotus Pond that met the tertiary drinking water standard were 100%, 74.91%, and 100% with great improvement, and the average concentrations of NH3-N, COD, and TP in Fushan Road Drainage have decreased by 91.43%, 10.49%, and 57.33% respectively, after the construction of the interception project. These indicated that the nitrogen and phosphorus pollution were successfully controlled by the control techniques in both locations, but COD concentration has to be addressed with more specialized strategies. Overall, the water quality improvement project for initial rainwater pollution plays a great role in effectively governing initial rainwater pollution and improving river water quality, and provides an effective technical reference for urban water ecological environment management.

Data on cloning, expression and biochemical characteristics of a chondroitin sulfate/dermatan sulfate 4-O-endosulfatase.

The data shown in this article are related to the published paper entitled "A novel 4-O-endosulfatase with high potential for the structure-function studies of chondroitin sulfate/dermatan sulfate" in Carbohydrate Polymers. In this article, the phylogenetic analysis, cloning, expression, purification, specificity and biochemical characteristics of the identified chondroitin sulfate/dermatan sulfate 4-O-endosulfatase (endoBI4SF) are described in detail. The recombinant endoBI4SF with a molecular mass of 59.13 kDa can can specifically hydrolyze the 4-O- but not 2-O- and 6-O-sulfate groups in the oligo-/polysaccharides of chondroitin sulfate/dermatan sulfate and show the maximum reaction rate in 50 mM Tris-HCl buffer (pH 7.0) at 50°C, which can be a very useful tool for the structural and functional studies of chondroitin sulfate/dermatan sulfate.

The role of ultrasonic cardiogram in the diagnosis of hypertension complicated with type 2 diabetes mellitus.

Hypertension diabetes mellitus is one of the serious complications of hypertension. In this study, ambulatory blood pressure monitoring (ABPM) and ultrasonic cardiogram (UCG) were used to investigate the cardiac changes and its influencing factors in hypertensive patients with type 2 diabetes mellitus. The ABPM, UCG, Hemoglobin A1c (HbA1c) and body mass index (BMI) of patients were examined. The comparison of HbA1c, BMI, gender, age, daytime and nighttime blood pressure, left ventricular mass index (LVMI), left ventricular hypertrophy (LVH), isovolumic relaxation time (IVRT) and E/A ratio were made between the two groups. The cardiac function of control group was better than that of group B, while that of group B was better than group A. The cardiac index level in group B was better than that in group A, but lower than that in control group. The LVMI in group A was clearly higher than group B and control group, and the incidence of LVH increased. In group A, the nocturnal systolic blood pressure was higher than control group and group B. Nocturnal diastolic blood pressure, daytime diastolic blood pressure and systolic blood pressure in groups A and B were higher than those the control group. The findings indicated that hypertension complicated with type 2 diabetes mellitus can cause degeneration of the heart, and complicated with type 2 diabetes mellitus can accelerate ventricular remodeling and functional deterioration. Hypertension with type 2 diabetes mellitus are more prone to left ventricular damage.

A novel 4-O-endosulfatase with high potential for the structure-function studies of chondroitin sulfate/dermatan sulfate.

The sulfation patterns of chondroitin sulfate (CS)/dermatan sulfate (DS), which encode unique biological information, play critical roles in the various biological functions of CS/DS chains. CS/DS sulfatases, which can specifically hydrolyze sulfate groups, could potentially be essential tools for deciphering and changing the biological information encoded by these sulfation patterns. However, endosulfatase with high activity to efficiently hydrolyze the sulfate groups inside CS/DS polysaccharides have rarely been identified, which hinders the practical applications of CS/DS sulfatases. Herein, a novel CS/DS 4-O-endosulfatase (endoBI4SF) with a strong ability to completely remove 4-O-sulfated groups inside various CS/DS polysaccharides was identified and successfully used to investigate the biological roles of 4-O-sulfated CS/DS in vitro and in vivo. This study provides a much-needed tool to tailor the sulfation patterns and explore the related functions of 4-O-sulfated CS/DS chains in vitro and in vivo.

Enzymatic Sequencing of Heparin Oligosaccharides Using Exolyase.

Heparin/heparan sulfate (HP/HS) is a class of acidic polysaccharides with many potential medical applications, especially HP, and its derivatives, low molecular weight heparins (LMWHs), have been widely used as anticoagulants to treat thrombosis for decades. However, the complex structure endows HP/HS a variety of biological functions and hinders the structural and functional studies of HP/HS. Heparinases derived from bacteria are useful tools for the structural studies of HP/HS as well as the preparation of LMWHs. The enzymatic method for the structural analysis of HP/HS chains is easy to operate, requires less samples, and is low cost. Here, we describe an enzymatic approach to investigate the primary sequences of the HP/HS oligosaccharides using a recently discovered exotype heparinase.

Predictive Value of Quantitative Duplex Ultrasound Analysis for In-stent Carotid Artery Restenosis.

Context: In-stent restenosis (ISR) is a common clinical complication after carotid artery stenting (CAS) and a major risk for a stent's fatigue life. Duplex ultrasound (DUS) is widely used for the preliminary evaluation and follow-up of extracranial carotid artery disease, but DUS stenosis grading is mainly based on the original or nonsurgical carotid artery. That grading may not be applicable to carotid artery stenosis after CAS. Objective: The study intended to investigate the predictive value of quantitative analysis of results from the DUS examination in the evaluation of ISR following CAS. Design: The research team designed a control analysis of result samples. Setting: The study took place in the Ultrasound Department at the Affiliated Yantai Yuhuangding Hospital of Qingdao University in Yantai, Shandong, China. Participants: Participants were 103 patients who underwent carotid artery stenting (CAS) between March 2017 and April 2018 at the hospital. Outcome Measures: The study used Doppler DUS and digital subtraction angiography (DSA) of the carotid artery at 12 months postoperatively to analyze the consistency of DUS and DSA in the evaluation of ISR. Taking the results of the DSA examination as the standard, the research team analyzed the differences between those results and the indicators from the DUS examination for participants with different severities of stenosis. The research team plotted the receiver operating characteristic curve (ROC) and evaluated the diagnostic efficiency of DUS indicators in the determination of restenosis, including diagnostic accuracy, sensitivity, specificity, positive predictive value, and negative predictive value. Results: The DSA examination showed that stenosis severity was 0%-30% for 51 participants, 31%-50% for 27 participants, 51%-80% for 16 participants, and >80% for 9 participants. The DUS showed that stenosis severity was 0%-30% for 35 participants, 31%-50% for 38 participants, 51%-80% for 22 participants, and >80% for 8 participants. The consistency was found to be Kappa (ĸ) = 0.74. Taking the DSA as the standard, the peak systolic velocity (PSV), end diastolic velocity (EDV), peak systolic velocity of the internal carotid artery/peak systolic velocity of the common carotid artery (PSVICA/PSVCCA) significantly increased in participants with a stenosis severity of 51-80% and >80%, compared with those with a stenosis severity of <50%, and the difference was statistically significant (P < .05). The ROC curve showed that the area under curve (AUC) of the PSV predicting restenosis at a >50% severity was significantly higher than those of the EDV and PSVICA/PSVCCA (P < .05). Where the optimal cut-off-off point for the PSV was 195 cm/s, the ROC curve showed that the AUC of the PSV predicting restenosis at an >80% severity was significantly higher than that of the EDV and PSVICA/PSVCCA (P < .05). Where the optimal cut-off point for the PSV was 280 cm/s, the PSV had significantly higher diagnostic accuracy, sensitivity, and positive predictive value than the EDV and PSVICA/PSVCCA in evaluating the restenosis at a severity of >50% and >80%. Conclusions: Doppler DUS can effectively evaluate restenosis after carotid artery stenting (CAS), where a PSV ≥195 cm/s and 280 cm/s can be used as the reference indicators for >50% and >80% restenosis.

A mutated glycosaminoglycan-binding domain functions as a novel probe to selectively target heparin-like epitopes on tumor cells.

The high heterogeneity and mutation rate of cancer cells often lead to the failure of targeted therapy, and therefore, new targets for multitarget therapy of tumors are urgently needed. Aberrantly expressed glycosaminoglycans (GAGs) have been shown to be involved in tumorigenesis and are promising new targets. Recently, the GAG-binding domain rVAR2 of the Plasmodium falciparum VAR2CSA protein was identified as a probe targeting cancer-associated chondroitin sulfate A-like epitopes. In this study, we found that rVAR2 could also bind to heparin (Hep) and chondroitin sulfate E. Therefore, we used rVAR2 as a model to establish a method based on random mutagenesis of the GAG-binding protein and phage display to identify and optimize probes targeting tumor GAGs. We identified a new probe, VAR2HP, which selectively recognized Hep by interacting with unique epitopes consisting of a decasaccharide structure that contains at least three HexA2S(1-4)GlcNS6S disaccharides. Moreover, we found that these Hep-like epitopes were overexpressed in various cancer cells. Most importantly, our in vivo experiments showed that VAR2HP had good biocompatibility and preferentially localizes to tumors, which indicates that VAR2HP has great application potential in tumor diagnosis and targeted therapy. In conclusion, this study provides a strategy for the discovery of novel tumor-associated GAG epitopes and their specific probes.

Ultra-light planar meta-absorber with wideband and full-polarization properties.

Absorbers have high potential application values in the military field, such as electronic screening, radar cross-section reduction and invisible cloaking. However, most methods have the defects of narrow bandwidth, low absorptivity, complex three-dimensional structure and fixed polarizations. In this paper, we realize an ultra-broadband and full-polarization planar metamaterial absorber (PMA) with a three-layer composite structure, which exhibits multi-resonant and impedance matching properties by combining the ultra-light foams and indium tin oxide (ITO) films. The bottom two layers achieve a high-efficiency absorption rate at the low and medium spectrum, while the upper layer realizes a absorption property at a high frequency. Also, an equivalent circuit model is extracted to explain its operating mechanism. The experimental results show that our meta-absorber can achieve great absorber performance of better than 90% within 1-18 GHz for full-polarization incident waves, which is in great agreement with the numerical simulations. Moreover, our device is insensitive to oblique incidences and polarizations and possesses the physical characteristics of an ultralight, weighing 0.6 kg for a square meter, which is only 1/85.0-1/126.7 of the conventional absorbers under the same size. All these excellent performances determine that our research can be a good candidate for military stealth materials.

Discovery of exolytic heparinases and their catalytic mechanism and potential application.

Heparinases (Hepases) are critical tools for the studies of highly heterogeneous heparin (HP)/heparan sulfate (HS). However, exolytic heparinases urgently needed for the sequencing of HP/HS chains remain undiscovered. Herein, a type of exolytic heparinases (exoHepases) is identified from the genomes of different bacteria. These exoHepases share almost no homology with known Hepases and prefer to digest HP rather than HS chains by sequentially releasing unsaturated disaccharides from their reducing ends. The structural study of an exoHepase (BIexoHep) shows that an N-terminal conserved DUF4962 superfamily domain is essential to the enzyme activities of these exoHepases, which is involved in the formation of a unique L-shaped catalytic cavity controlling the sequential digestion of substrates through electrostatic interactions. Further, several HP octasaccharides have been preliminarily sequenced by using BIexoHep. Overall, this study fills the research gap of exoHepases and provides urgently needed tools for the structural and functional studies of HP/HS chains.

Identification and Action Patterns of Two Chondroitin Sulfate Sulfatases From a Marine Bacterium Photobacterium sp. QA16.

Chondroitin sulfate (CS)/dermatan sulfate (DS) is a kind of sulfated polyanionic, linear polysaccharide belonging to glycosaminoglycan. CS/DS sulfatases, which specifically hydrolyze sulfate groups from CS/DS oligo-/polysaccharides, are potential tools for structural and functional studies of CD/DS. However, only a few sulfatases have been reported and characterized in detail to date. In this study, two CS/DS sulfatases, PB_3262 and PB_3285, were identified from the marine bacterium Photobacterium sp. QA16 and their action patterns were studied in detail. PB_3262 was characterized as a novel 4-O-endosulfatase that can effectively and specifically hydrolyze the 4-O-sulfate group of disaccharide GlcUAß1-3GalNAc(4-O-sulfate) but not GlcUAß1-3GalNAc(4,6-O-sulfate) and IdoUAα1-3GalNAc(4-O-sulfate) in CS/DS oligo-/polysaccharides, which is very different from the identified 4-O-endosulfatases in the substrate profile. In contrast, PB_3285 specifically hydrolyzes the 6-O-sulfate groups of GalNAc(6-O-sulfate) residues located at the reducing ends of the CS chains and is the first recombinantly expressed 6-O-exosulfatase to effectively act on CS oligosaccharides.

The Bonding Mechanism of the Micro-Interface of Polymer Coated Steel.

As food and beverages require more and more green and safe packaging products, the emergence of polymer coated steel (PCS) has been promoted. PCS is a layered composite strip made of metal and polymer. To probe the bonding mechanism of PCS micro-interface, the substrate tin-free steel (TFS) was physically characterized by SEM and XPS, and cladding polyethylene terephthalate (PET) was simulated by first-principles methods of quantum mechanics (QM). We used COMPASS force field for molecular dynamics (MD) simulation. XPS pointed out that the element composition of TFS surface coating is Cr(OH)3, Cr2O3 and CrO3. The calculation results of MD and QM indicate that the chromium oxide and PET molecules compound in the form of acid-base interaction. The binding energies of Cr2O3 (110), (200), and (211) with PET molecules are -13.07 eV, -2.74 eV, and -2.37 eV, respectively. We established a Cr2O3 (200) model with different hydroxyl concentrations. It is proposed that the oxygen atom in C=O in the PET molecule combines with -OH on the surface of TFS to form a hydrogen bond. The binding energy of the PCS interface increases with the increase of the surface hydroxyl concentration of the TFS. It provides theoretical guidance and reference significance for the research on the bonding mechanism of PCS.

Identification and biochemical characterization of a novel chondroitin sulfate/dermantan sulfate lyase from Photobacterium sp.

Chondroitin sulfate (CS)/dermatan sulfate (DS) lyases play important roles in structural and functional studies of CS/DS. In this study, a novel CS/DS lyase (enCSase) was identified from the genome of the marine bacterium Photobacterium sp. QA16. This enzyme is easily heterologously expressed and purified as highly active form against various CS, DS and hyaluronic acid (HA). Under the optimal conditions, the specific activities of this enzyme towards CSA, CSC, CSD, CSE, DS and HA were 373, 474, 171, 172, 141 and 97 U/mg of proteins, respectively. As an endolytic enzyme, enCSase degrades HA to unsaturated hexa- and tetrasaccharides but CS/DS to unsaturated tetra- and disaccharides as the final products. Sequencing analysis showed that the structures of tetrasaccharides in the final products of CS variants were not unique but were highly variable, indicating the randomness of substrate degradation by this enzyme. Further studies showed that the smallest substrate of enCSase was octasaccharide for HA but hexasaccharide for CS/DS, which could explain why this enzyme cannot degrade HA hexa- and tetrasaccharides and CS/DS tetrasaccharides further. It is believed that enCSase may be a very useful tool for structural and functional studies and related applications of CS/DS and HA.

Biochemical characteristics and synergistic effect of two novel alginate lyases from Photobacterium sp. FC615.

BACKGROUND: Macroalgae and microalgae, as feedstocks for third-generation biofuel, possess competitive strengths in terms of cost, technology and economics. The most important compound in brown macroalgae is alginate, and the synergistic effect of endolytic and exolytic alginate lyases plays a crucial role in the saccharification process of transforming alginate into biofuel. However, there are few studies on the synergistic effect of endolytic and exolytic alginate lyases, especially those from the same bacterial strain. RESULTS: In this study, the endolytic alginate lyase AlyPB1 and exolytic alginate lyase AlyPB2 were identified from the marine bacterium Photobacterium sp. FC615. These two enzymes showed quite different and novel enzymatic properties whereas behaved a strong synergistic effect on the saccharification of alginate. Compared to that when AlyPB2 was used alone, the conversion rate of alginate polysaccharides to unsaturated monosaccharides when AlyPB1 and AlyPB2 acted on alginate together was dramatically increased approximately sevenfold. Furthermore, we found that AlyPB1 and AlyPB2 acted the synergistic effect basing on the complementarity of their substrate degradation patterns, particularly due to their M-/G-preference and substrate-size dependence. In addition, a novel method for sequencing alginate oligosaccharides was developed for the first time by combining the 1H NMR spectroscopy and the enzymatic digestion with the exo-lyase AlyPB2, and this method is much simpler than traditional methods based on one- and two-dimensional NMR spectroscopy. Using this strategy, the sequences of the final tetrasaccharide and pentasaccharide product fractions produced by AlyPB1 were easily determined: the tetrasaccharide fractions contained two structures, ΔGMM and ΔMMM, at a molar ratio of 1:3.2, and the pentasaccharide fractions contained four structures, ΔMMMM, ΔMGMM, ΔGMMM, and ΔGGMM, at a molar ratio of ~ 1:1.5:3.5:5.25. CONCLUSIONS: The identification of these two novel alginate lyases provides not only excellent candidate tool-type enzymes for oligosaccharide preparation but also a good model for studying the synergistic digestion and saccharification of alginate in biofuel production. The novel method for oligosaccharide sequencing described in this study will offer a very useful approach for structural and functional studies on alginate.

Comparative Study of Two Chondroitin Sulfate/Dermatan Sulfate 4-O-Sulfatases With High Identity.

Chondroitin sulfate/dermatan sulfate (CS/DS) sulfatases are potential tools for structural and functional studies of CD/DS chains. In our previous study, a CS/DS 4-O-endosulfatase (endoVB4SF) was identified from a marine bacterium (Wang et al., 2015). Herein, another CS/DS 4-O-sulfatase (exoPB4SF) was identified from a Photobacterium sp. ExoPB4SF shares an 83% identity with endoVB4SF but showed strict exolytic activity. Comparative studies were performed for both enzymes on the basis of biochemical features, substrate-degrading patterns and three-dimensional structures. exoPB4SF exhibited a wider temperature and pH adaptability and better thermostability than endoVB4SF. Furthermore, exoPB4SF is a strict exolytic sulfatase that only releases the sulfate group from the GalNAc residue located at the reducing end, whereas endoVB4SF preferentially removed sulfate esters from the reducing end toward the non-reducing end though its directional degradation property was not strict. In addition, the structure of endoVB4SF was determined by X-ray crystallography at 1.95 Å. It adopts a globular conformation with two monomers per asymmetric unit. The exoPB4SF structure was constructed by homology modeling. Molecular docking results showed that although the residues around the catalytic center are conserved, the residues at the active site of endoVB4SF adopted a more favorable conformation for the binding of long CS/DS chains than those of exoPB4SF, which may explain why the two highly homogenous sulfatases possessed different action patterns. The results of this study provide insight into the structure-function relationship of CS/DS endo- and exosulfatases for the first time.