Hyaluronic acid production by recombinant Lactococcus lactis

Chien, Liang-Jung; Lee, Cheng‐Kang

doi:10.1007/s00253-007-1153-z

Cited by 105 publications

(78 citation statements)

References 19 publications

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“…As expected, an undersupply of UDPGlcA and UDP-GlcNAc decreased the yield. From these results, we can conclude that HasA is essential for HA production in L. lactis and improving the precursor sugar biosynthesis levels can increase the production of HA, confirming previous findings [4].…”

Section: Food-grade Ha Production In the Recombinant Strainssupporting

confidence: 91%

“…Several studies also showed that expressing endogenous UDP-GlcDH, which directs a step in the synthesis pathway of UDP-GlcA, could improve HA production [4,17,26]. We hypothesized that the endogenous enzymes involved in the synthesis of the UDP sugars should work more efficiently than the heterologous enzymes from S. zooepidemicus in L. lactis.…”

Section: Food-grade Ha Production In the Recombinant Strainsmentioning

confidence: 95%

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Constructing a recombinant hyaluronic acid biosynthesis operon and producing food-grade hyaluronic acid in Lactococcus lactis

Sheng

Ling

Wang

2015

Journal of Industrial Microbiology and Biotechnology

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Hyaluronic acid (HA), a natural high molecular weight polysaccharide, is produced by Streptococcus zooepidemicus. However, Streptococcus has several drawbacks including its potential to produce exotoxins, so there is demand for an alternative HA source. Here, a recombinant HA biosynthesis operon, as well as the HA biosynthesis operon of S. zooepidemicus were introduced into L. lactis using the nisin-controlled expression system, respectively. HA was successfully synthesized by recombinant L. lactis. Furthermore, overexpression of the endogenous enzymes directing the synthesis of precursor sugars was effective at increasing HA production, and increasing the supply of UDP-activated monosaccharide donors aided synthesis of monodisperse HA polysaccharides. Besides GRAS host strain (L. lactis) and NICE system, the selecting marker (lacF gene) of the recombinant strain is also food grade. Therefore, HA produced by recombinant L. lactis overcomes the problems associated with Streptococcus and provides a source of food-grading HA appropriate for widespread biotechnological applications.

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Section: Food-grade Ha Production In the Recombinant Strainssupporting

confidence: 91%

Section: Food-grade Ha Production In the Recombinant Strainsmentioning

confidence: 95%

Constructing a recombinant hyaluronic acid biosynthesis operon and producing food-grade hyaluronic acid in Lactococcus lactis

Sheng

Ling

Wang

2015

Journal of Industrial Microbiology and Biotechnology

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“…HA has been produced in heterologous hosts, including Enterococcus faecalis (45), Escherichia coli (9,38), B. subtilis (46), and L. lactis (47). Given superior tools for strain engineering of the latter four hosts, it may prove easier to engineer microbial high molecular weight HA producers by targeting the UDP-GlcNAc/UDP-GlcUA ratio in these hosts.…”

Section: Discussionmentioning

confidence: 99%

Hyaluronan Molecular Weight Is Controlled by UDP-N-acetylglucosamine Concentration in Streptococcus zooepidemicus

Chen

Marcellin

Hung

et al. 2009

Journal of Biological Chemistry

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The molecular weight of hyaluronan is important for its rheological and biological function. The molecular mechanisms underlying chain termination and hence molecular weight control remain poorly understood, not only for hyaluronan synthases but also for other ␤-polysaccharide synthases, e.g. cellulose, chitin, and 1,3-betaglucan synthases. In this work, we manipulated metabolite concentrations in the hyaluronan pathway by overexpressing the five genes of the hyaluronan synthesis operon in Streptococcus equi subsp. zooepidemicus. Overexpression of genes involved in UDP-glucuronic acid biosynthesis decreased molecular weight, whereas overexpression of genes involved in UDP-N-acetylglucosamine biosynthesis increased molecular weight. The highest molecular mass observed was at 3.4 ؎ 0.1 MDa twice that observed in the wild-type strain, 1.8 ؎ 0.1 MDa. The data indicate that (a) high molecular weight is achieved when an appropriate balance of UDP-N-acetylglucosamine and UDP-glucuronic acid is achieved, (b) UDP-N-acetylglucosamine exerts the dominant effect on molecular weight, and (c) the wild-type strain has suboptimal levels of UDP-Nacetylglucosamine. Consistent herewith molecular weight correlated strongly ( ‫؍‬ 0.84, p ‫؍‬ 3 ؋ 10 ؊5 ) with the concentration of UDP-N-acetylglucosamine. Data presented in this paper represent the first model for hyaluronan molecular weight control based on the concentration of activated sugar precursors. These results can be used to engineer strains producing high molecular weight hyaluronan and may provide insight into similar polymerization mechanisms in other polysaccharides.

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“…A produção de ácido hialurônico por Streptococcus é bastante conhecida, estando presente nas cápsulas das espécies Streptococcus thermophilus (IZAWA et al, 2009), Streptococcus pyogenes (KANG et al, 2012) e Streptococcus zooepidemicus (PATIL et al, 2011), além de Chlorella e Pasteurella multocida (DeANGELIS, 2012). Entretanto, a inserção de genes has que codificam para a enzima hialurona sintase, responsável por sua síntese, também foi eficiente para Bacillus subtilis (WIDNER et al, 2005) e Lactococcus lactis (CHIEN;LEE, 2007) produzirem esse polímero.…”

Section: Seleção De Bactérias Com Características De Rpcpunclassified

Bactérias associadas às cactáceas da Caatinga: promoção de crescimento de plantas sob estresse hídrico

Kavamura¹

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AGRADECIMENTOS Silva, Cassiano Forner, José Abrahão Haddad Galvão, Leonardo José da Silva (Léo), Lúcio Bertoldo Costa, Michelli de Souza dos Santos, Milena Duarte Lançoni, Rafael Eduardo Silva, Regiane Iost (Figura) A Caatinga, bioma exclusivamente brasileiro, inserido no clima semiárido nordestino, apresenta xerófitas com alta resistência aos períodos de seca. Estas plantas associam-se a micro-organismos que também se encontram bem adaptados, desenvolvendo mecanismos de proteção celular contra o estresse hídrico, assim como proteção vegetal contra os efeitos negativos da dessecação. O presente estudo buscou compreender as bactérias associadas às cactáceas da Caatinga, analisando a estrutura das comunidades bacterianas de solo e da rizosfera de Cereus jamacaru durante a alteração do período chuvoso para o de seca, identificando os grupos dominantes e discutindo algumas funções que possibilitem a manutenção da interação solo-cacto-micro-organismo durante o período de seca. Além disso, buscou selecionar bactérias tolerantes à seca e que fossem capazes de promover crescimento de plantas sob estresse hídrico. Amostras foram coletadas ao longo da Caatinga, em cinco estados: BA, CE, PI, PB e RN totalizando cinco pontos. Com o uso de metodologias independentes de cultivo, foi possível observar que o período de amostragem (chuvoso/seca) foi o principal responsável pela alteração na estrutura das comunidades bacterianas. Os filos Proteobacteria e Bacteroidetes foram abundantes durante o período chuvoso e os filos Actinobacteria e o gênero Bacillus abundantes durante o período de seca. Com o uso de metodologias dependentes de cultivo, foram isoladas com bastante frequência linhagens pertencentes ao gênero Bacillus, capazes de crescer em meio com reduzida atividade de água e com alguns mecanismos de proteção contra a dessecação, como a produção de exopolissacarídeos e biofilme. Além disso, várias linhagens apresentaram mecanismos de promoção de crescimento de plantas diretos e/ou indiretos, como produção de fitohormônio, disponibilização de P por meio de solubilização, fixação de nitrogênio, redução dos efeitos negativos do estresse causados por etileno, produção de celulase e amônia. Uma linhagem de Bacillus sp. foi capaz de promover crescimento de milho sob estresse hídrico, incrementando alguns parâmetros vegetais analisados. São discutidos o uso de consórcio bacteriano entre duas linhagens, além dos mecanismos que os micro-organismos dispõem para tolerar condições ambientais adversas e ainda as funções que estes micro-organismos podem ter na proteção vegetal contra a dessecação. Caatinga, a Brazilian unique biome, inserted in the semi-arid climate of Brazilian northeast, presents xerophytes with high resistance to drought periods. These plants are found in association with well-adapted microorganisms that have evolved some cellular protection mechanisms against water stress, as well as plant protection against the negative effects of desiccation. This work aimed to study the bacteria associated to some Caatinga cact...

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Hyaluronic acid production by recombinant Lactococcus lactis

Cited by 105 publications

References 19 publications

Constructing a recombinant hyaluronic acid biosynthesis operon and producing food-grade hyaluronic acid in Lactococcus lactis

Constructing a recombinant hyaluronic acid biosynthesis operon and producing food-grade hyaluronic acid in Lactococcus lactis

Hyaluronan Molecular Weight Is Controlled by UDP-N-acetylglucosamine Concentration in Streptococcus zooepidemicus

Bactérias associadas às cactáceas da Caatinga: promoção de crescimento de plantas sob estresse hídrico

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