Produção e purificação da proteína recombinante humana SLPI a partir da microalga Chlamydomonas reinhardtii
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Data
2023-02-28
Autores
Silva, Bárbara Luísa Chagas da
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Universidade Estadual Paulista (Unesp)
Resumo
O inibidor de protease secretado por leucócitos (SLPI) foi primeiramente descrito em 1986. Desde então, é alvo de diversos estudos devido às suas variadas funções. A proteína SLPI desempenha diversos papeis no organismo humano, sendo que a principal é sua ação inibidora de serina – proteases. Devido à essa função, a SLPI têm um importante papel na manutenção da homeostase entre proteases e antiproteases no trato respiratório humano e na pele, agindo principalmente em respostas anti-inflamatórias contra agentes danosos ao tecido. Adicionalmente, a proteína SLPI também já foi caracterizada em mecanismos de cicatrização, metabolismo ósseo, câncer e apresenta propriedades antimicrobianas, antifúngicas e antivirais. Por ser uma proteína multifuncional, o potêncial terapeutico da SLPI é bastante explorado por pesquisadores e vem sendo testado em diferentes campos da ciência, como doenças respiratórias, neuroregeneração, cicatrização de feridas, e cardioproteção em situações de isquemia/reperfusão. Devido ao crescente interesse em pesquisas biotecnológicas visando a proteína SLPI como um bioproduto, diversas empresas passaram a produzir e comercializar a proteína em diferentes sistemas de expressão e produção. Porém, a SLPI ainda é vendida somente para fins de pesquisa científica à um custo muito alto. O mercado biotecnológico de produção e expessão de proteínas recombinantes têm avançado bastante nos últimos anos, e diversas plataformas estão sendo desenvolvidas, como plantas, animais transgênicos, células humanas e microalgas. As microalgas vem sendo muito utilizadas em pesquisas para biorremediação, desenvolvimento de ração e suplemento alimentar, e não há muito tempo, ganharam notável evidência como promissoras plataformas para produção de proteínas recombinantes. Esses organismos apresentam grande potencial como plataformas biotecnológicas para desenvolvimento de bioprodutos, pois são seres eucariotos, unicelulares, de rápido crescimento e com baixo custo de meio de cultura. À vista disso, o presente projeto teve como objetivo a produção e purificação da proteína recombinante humana SLPI a partir da microalga Chlamydomonas reinhardtii.
The secretory leukocyte protease inhibitor (SLPI) was first described in 1986. Since then, it has been the subject of several studies due to its varied functions. The SLPI protein plays several roles in the human body, the main one being its inhibitory action on serine-proteases. Due to this function, SLPI play an important role in maintaining homeostasis between proteases and antiproteases in the human respiratory tract and in the skin, acting mainly in anti-inflammatory responses against tissue-damaging agents. Additionally, the SLPI protein has also been characterized in healing mechanisms, bone metabolism, cancer and has antimicrobial, antifungal and antiviral properties. As it is a multifunctional protein, the therapeutic potential of SLPI is widely explored by researchers and has been tested in different fields of science, such as respiratory diseases, neuroregeneration, wound healing, and cardioprotection in situations of ischemia/reperfusion. Due to the growing interest in biotechnological research, targeting the SLPI protein as a by-product, several companies started to produce and commercialize the protein in different expression and production systems. However, SLPI is still only sold for scientific research purposes at a very high cost. The biotechnological market for the production and expression of recombinant proteins has advanced a lot in recent years, and several platforms are being developed, such as plants, transgenic animals, human cells and microalgae. Microalgae have been widely used in research for bioremediation, feed development and food supplement, and not long ago, they gained the spotlight as promising platforms for the production of recombinant proteins. These organisms have great potential as biotechnological platforms for the development of bioproducts, as they are eukaryotic, unicellular, fast-growing beings with low cost of culture medium. In view of this, the present project aimed at the production and purification of the recombinant human protein SLPI from the microalgae Chlamydomonas reinhardtii.
The secretory leukocyte protease inhibitor (SLPI) was first described in 1986. Since then, it has been the subject of several studies due to its varied functions. The SLPI protein plays several roles in the human body, the main one being its inhibitory action on serine-proteases. Due to this function, SLPI play an important role in maintaining homeostasis between proteases and antiproteases in the human respiratory tract and in the skin, acting mainly in anti-inflammatory responses against tissue-damaging agents. Additionally, the SLPI protein has also been characterized in healing mechanisms, bone metabolism, cancer and has antimicrobial, antifungal and antiviral properties. As it is a multifunctional protein, the therapeutic potential of SLPI is widely explored by researchers and has been tested in different fields of science, such as respiratory diseases, neuroregeneration, wound healing, and cardioprotection in situations of ischemia/reperfusion. Due to the growing interest in biotechnological research, targeting the SLPI protein as a by-product, several companies started to produce and commercialize the protein in different expression and production systems. However, SLPI is still only sold for scientific research purposes at a very high cost. The biotechnological market for the production and expression of recombinant proteins has advanced a lot in recent years, and several platforms are being developed, such as plants, transgenic animals, human cells and microalgae. Microalgae have been widely used in research for bioremediation, feed development and food supplement, and not long ago, they gained the spotlight as promising platforms for the production of recombinant proteins. These organisms have great potential as biotechnological platforms for the development of bioproducts, as they are eukaryotic, unicellular, fast-growing beings with low cost of culture medium. In view of this, the present project aimed at the production and purification of the recombinant human protein SLPI from the microalgae Chlamydomonas reinhardtii.
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Palavras-chave
Biotecnologia, Microalga, Proteínas recombinantes, SLPI, Chlamydomonas reinhardtii, Purificação, Purification, Recombinant protein