Production of recombinant human N-acetylgalactosamine-6-sulfate sulfatase enzyme in Pichia pastoris

Rodríguez, Arturo; Moreno, Jefferson; Sánchez, Jhonnathan; Suescún-Díaz, Daniel; Beltran, Laura; Espejo, Ángela J.; Almeciga, Carlos; Barrera, Luis Alejandro

doi:10.1016/j.ymgme.2012.11.211

Cited by 3 publications

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“…Nevertheless, as mentioned before the activities are not comparable between microorganisms systems and CHO cells due to substantial differences in the enzyme activity assays. However, western-blot analysis showed that the recombinant enzyme has a similar band profile to the human wild-type GALNS from leucocytes, as well as similar temperature and pH stability profiles to those reported for the recombinant enzyme produced in CHO cells [65]. It is noteworthy that the recombinant GALNS produced in P. pastoris was captured by HEK293 cells and Morquio A skin fibroblast, showing the potential of this enzyme towards the development of an ERT for MPS IV A.…”

Section: Recombinant N-acetylgalactosamine-6-sulfate Sulfatasesupporting

confidence: 62%

“…Recently ERT for Morquio A disease was approved, which showed a modest improvement in a 6-min walk test (6MWT), and a reduction of urinary KS, after a 24-weeks treatment [62]. As an alternative, recombinant GALNS have been produced in E. coli K12 [11,63,64] and in the methylotrophic yeast P. pastoris [65,66] (Table 1).…”

Section: Recombinant N-acetylgalactosamine-6-sulfate Sulfatasementioning

confidence: 99%

“…(Fig. 4) [65,66]. Human GALNS cDNA, with the native or alpha-factor signal peptide, was inserted into P. pastoris GS115 under the control of the alcohol oxidase 1 gene promoter.…”

Section: Recombinant N-acetylgalactosamine-6-sulfate Sulfatasementioning

confidence: 99%

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Human recombinant lysosomal enzymes produced in microorganisms

Espejo-Mojica

Alméciga-Díaz

Rodríguez

et al. 2015

Molecular Genetics and Metabolism

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Section: Recombinant N-acetylgalactosamine-6-sulfate Sulfatasesupporting

confidence: 62%

Section: Recombinant N-acetylgalactosamine-6-sulfate Sulfatasementioning

confidence: 99%

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Human recombinant lysosomal enzymes produced in microorganisms

Espejo-Mojica

Alméciga-Díaz

Rodríguez

et al. 2015

Molecular Genetics and Metabolism

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“…Recombinant GALNS has been produced in Chinese hamster ovary (CHO) cells, with enzyme activities of approximately 170,000 nmol/h/mg 78 and 120,000 nmol/h/mg; 79 however, these activity values are not equivalent because they have been obtained using substantially different methods. Production of recombinant GALNS in E. coli BL21 (DE3) (erGALNS) 32 , 80 , 81 and in the methylotrophic yeast Pichia pastoris (prGALNS) 33 , 82 has emerged as an alternative to conventional production in CHO cells. In E. coli , as well as in other prokaryotes, cysteine- and serine-sulfatase types have been identified, which are activated by a formylglycine-generating enzyme and anaerobic sulfatase-maturating enzyme (AtsB), respectively, in these microorganisms.…”

Section: Enzyme Replacement Therapymentioning

confidence: 99%

“…These limitations are also observed for ERT for other forms of MPS. 7 , 27 – 29 To address the above issues, an improved long circulating ERT and a bone-targeting enzyme are proposed, 30 , 31 as well as the use of microorganism for the production of recombinant GALNS, 32 , 33 which could potentially help to lower the cost.…”

Section: Introductionmentioning

confidence: 99%

Impact of enzyme replacement therapy and hematopoietic stem cell transplantation in patients with Morquio A syndrome

Tomatsu¹,

Sawamoto²,

Alméciga-Díaz³

et al. 2015

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Patients with mucopolysaccharidosis IVA (MPS IVA) can present with systemic skeletal dysplasia, leading to a need for multiple orthopedic surgical procedures, and often become wheelchair bound in their teenage years. Studies on patients with MPS IVA treated by enzyme replacement therapy (ERT) showed a sharp reduction on urinary keratan sulfate, but only modest improvement based on a 6-minute walk test and no significant improvement on a 3-minute climb-up test and lung function test compared with the placebo group, at least in the short-term. Surgical remnants from ERT-treated patients did not show reduction of storage materials in chondrocytes. The impact of ERT on bone lesions in patients with MPS IVA remains limited. ERT seems to be enhanced in a mouse model of MPS IVA by a novel form of the enzyme tagged with a bone-targeting moiety. The tagged enzyme remained in the circulation much longer than untagged native enzyme and was delivered to and retained in bone. Three-month-old MPS IVA mice treated with 23 weekly infusions of tagged enzyme showed marked clearance of the storage materials in bone, bone marrow, and heart valves. When treatment was initiated at birth, reduction of storage materials in tissues was even greater. These findings indicate that specific targeting of the enzyme to bone at an early stage may improve efficacy of ERT for MPS IVA. Recombinant N-acetylgalactosamine-6-sulfate sulfatase (GALNS) in Escherichia coli BL21 (DE3) (erGALNS) and in the methylotrophic yeast Pichia pastoris (prGALNS) has been produced as an alternative to the conventional production in Chinese hamster ovary cells. Recombinant GALNS produced in microorganisms may help to reduce the high cost of ERT and the introduction of modifications to enhance targeting. Although only a limited number of patients with MPS IVA have been treated with hematopoietic stem cell transplantation (HSCT), beneficial effects have been reported. A wheelchair-bound patient with a severe form of MPS IVA was treated with HSCT at 15 years of age and followed up for 10 years. Radiographs showed that the figures of major and minor trochanter appeared. Loud snoring and apnea disappeared. In all, 1 year after bone marrow transplantation, bone mineral density at L2–L4 was increased from 0.372 g/cm2 to 0.548 g/cm2 and was maintained at a level of 0.48±0.054 for the following 9 years. Pulmonary vital capacity increased approximately 20% from a baseline of 1.08 L to around 1.31 L over the first 2 years and was maintained thereafter. Activity of daily living was improved similar to the normal control group. After bilateral osteotomies, a patient can walk over 400 m using hip–knee–ankle–foot orthoses. This long-term observation of a patient shows that this treatment can produce clinical improvements although bone deformity remained unchanged. In conclusion, ERT is a therapeutic option for MPS IVA patients, and there are some indications that HSCT may be an alternative to treat this disease. However, as neither seems to be a curative therapy, at least for the sk...

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Mucopolysaccharidosis IVA and glycosaminoglycans

Khan

Alméciga-Díaz

Sawamoto

et al. 2017

Molecular Genetics and Metabolism

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Mucopolysaccharidosis IVA (MPS IVA; Morquio A: OMIM 253000) is a lysosomal storage disease with an autosomal recessive trait caused by the deficiency of N-acetylgalactosamine-6-sulfate sulfatase. Deficiency of this enzyme leads to accumulation of specific glycosaminoglycans (GAGs): chondroitin-6-sulfate (C6S) and keratan sulfate (KS). C6S and KS are mainly produced in the cartilage. Therefore, the undegraded substrates are stored primarily in cartilage and in its extracellular matrix (ECM), leading to a direct impact on cartilage and bone development, and successive systemic skeletal dysplasia. Chondrogenesis, the earliest phase of skeletal formation, is maintained by cellular interactions with the ECM, growth and differentiation factors, signaling pathways, and transcription factors in a temporal-spatial manner. In patients with MPS IVA, the cartilage is disrupted at birth as a consequence of abnormal chondrogenesis and/or endochondral ossification. The unique skeletal features are distinguished by a disproportional short stature, odontoid hypoplasia, spinal cord compression, tracheal obstruction, pectus carinatum, kyphoscoliosis, platyspondyly, coxa valga, genu valgum, waddling gait, and laxity of joints. In spite of many descriptions of these unique clinical features, delay of diagnosis still happens. The pathogenesis and treatment of systemic skeletal dysplasia in MPS IVA remains an unmet challenge. In this review article, we comprehensively describe historical aspect, property of GAGs, diagnosis, screening, pathogenesis, and current and future therapies of MPS IVA.

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Production of recombinant human N-acetylgalactosamine-6-sulfate sulfatase enzyme in Pichia pastoris

Cited by 3 publications

References 0 publications

Human recombinant lysosomal enzymes produced in microorganisms

Human recombinant lysosomal enzymes produced in microorganisms

Impact of enzyme replacement therapy and hematopoietic stem cell transplantation in patients with Morquio A syndrome

Mucopolysaccharidosis IVA and glycosaminoglycans

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Production of recombinant human N-acetylgalactosamine-6-sulfate sulfatase enzyme in Pichia pastoris

Cited by 3 publications

References 0 publications

Human recombinant lysosomal enzymes produced in microorganisms

Human recombinant lysosomal enzymes produced in microorganisms

Impact of enzyme replacement therapy and&nbsp;hematopoietic stem cell transplantation in&nbsp;patients with Morquio A syndrome

Mucopolysaccharidosis IVA and glycosaminoglycans

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Product

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Impact of enzyme replacement therapy and hematopoietic stem cell transplantation in patients with Morquio A syndrome