Entrapment of phenylalanine ammonia-lyase in silk fibroin for protection from proteolytic attack |
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| Authors: | S Inoue Y Matsunaga H Iwane M Sotomura T Nose |
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| Institution: | 2. Laboratório de Pesquisas em Leishmanioses, Instituto de Doenças Tropicais Natan Portela–IDTNP, Teresina 64001450, Brazil;3. EMBRAPA Recursos Genéticos e Biotecnologia, 70770-917 Brasília, DF, Brazil;4. Laboratório de Espectrometria de Massa, LEM, Sala de Nanotecnologia, EMBRAPA, Recursos Genéticos e Biotecnologia, Brasília, DF 70770-917, Brazil;5. Laboratório Interdisciplinar de Materiais Avançados, LIMAV, CCN, UFPI, Teresina, PI 64049-550, Brazil;1. Institute of Complex Systems, Structural Biochemistry (ICS-6), Research Center Jülich, 52428 Jülich, Germany;2. Central Institute for Engineering, Electronics and Analytics (ZEA-3), Research Center Jülich, 52428 Jülich, Germany;3. Institut für Physikalische Biologie, Heinrich-Heine-Universität Düsseldorf, 40225 Düsseldorf, Germany |
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| Abstract: | Phenylalanine ammonia-lyase was entrapped in silk fibroin. The entrapped enzyme showed a similar Km for Phe and pH optimum to the free enzyme. It was resistant against chymotrypsin and trypsin in vitro. To assess the activity in vivo, the free or entrapped enzymes and then Phe were injected into rat duodenum, and cinnamate, a product, in plasma was determined as the most direct evidence of the enzyme activity. The entrapped enzyme but not the free form caused a marked raise of plasma cinnamate. It declined with a half life of about 45 min, which was significantly longer than that (10-15 min) observed upon i.v. administration of cinnamate. These results indicated that the entrapped enzyme was actively degrading Phe in the intestinal tract. Entrapment of phenylalanine ammonia-lyase in fibroin thus provides a new prospect for oral enzyme therapy of phenylketonuria. |
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