Abstract:
BACKGROUND:Lipases (EC 3.1.1.3) catalyze the hydrolysis of triacyl glycerol to glycerol and are involved in the synthesis of both short chain and long chain acylglycerols. They are widely used industrially in various applications, such as baking, laundry detergents and as biocatalysts in alternative energy strategies. Marine ecosystems are known to represent a large reservoir of biodiversity with respect to industrially useful enzymes. However the vast majority of microorganisms within these ecosystems are not readily culturable. Functional metagenomic based approaches provide a solution to this problem by facilitating the identification of novel enzymes such as the halo-tolerant lipase identified in this study from a marine sponge metagenome. RESULTS:A metagenomic library was constructed from the marine sponge Haliclona simulans in the pCC1fos vector, containing approximately 48,000 fosmid clones. High throughput plate screening on 1% tributyrin agar resulted in the identification of 58 positive lipase clones. Following sequence analysis of the 10 most highly active fosmid clones the pCC1fos53E1 clone was found to contain a putative lipase gene lpc53E1, encoded by 387 amino acids and with a predicted molecular mass of 41.87 kDa. Sequence analysis of the predicted amino acid sequence of Lpc53E1 revealed that it is a member of the group VIII family of lipases possessing the SXTK motif, related to type C β-lactamases. Heterologous expression of lpc53E1 in E. coli and the subsequent biochemical characterization of the recombinant protein, showed an enzyme with the highest substrate specificity for long chain fatty acyl esters. Optimal activity was observed with p- nitrophenyl palmitate (C16) at 40°C, in the presence of 5 M NaCl at pH 7; while in addition the recombinant enzyme displayed activity across broad pH (3-12) and temperature (4 -60°C) ranges and high levels of stability in the presence of various solvents at NaCl concentrations as high as 5 M and at temperatures ranging from 10 to 80°C. A maximum lipase activity of 2,700 U/mg was observed with 10 mM p-nitrophenyl palmitate as substrate, in the presence of 5 mM Ca2+ and 5 M NaCl, and a reaction time of 15 min at pH 7 and 40°C; while KM and Vmax values were calculated to be 1.093 mM-1 and 50 μmol/min, respectively. CONCLUSION:We have isolated a novel halo tolerant lipase following a functional screen of a marine sponge fosmid metagenomic library. The activity and stability profile of the recombinant enzyme over a wide range of salinity, pH and temperature; and in the presence of organic solvent and metal ions suggests a utility for this enzyme in a variety of industrial applications.
journal_name
Microb Cell Factjournal_title
Microbial cell factoriesauthors
Selvin J,Kennedy J,Lejon DP,Kiran GS,Dobson ADdoi
10.1186/1475-2859-11-72subject
Has Abstractpub_date
2012-06-01 00:00:00pages
72issn
1475-2859pii
1475-2859-11-72journal_volume
11pub_type
杂志文章abstract:BACKGROUND:Probiotic bacteria are increasingly used as immunomodulatory agents. Yet detailed molecular knowledge on the immunomodulatory molecules of these bacteria is lagging behind. Lipoteichoic acid (LTA) is considered a major microbe-associated molecular pattern (MAMP) of Gram-positive bacteria. However, many detai...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-11-161
更新日期:2012-12-15 00:00:00
abstract:BACKGROUND:Hepatitis C virus (HCV) is a common and leading cause for liver cirrhosis and hepatocellular carcinoma. Current therapies to treat HCV infection are shown to be partially effective and poorly tolerated. Therefore, ample efforts are underway to rationally design therapies targeting the HCV non-structural prot...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-12-111
更新日期:2013-11-16 00:00:00
abstract:BACKGROUND:cis, cis-Muconic acid is an important chemical that can be biosynthesized from simple substrates in engineered microorganisms. Recently, it has been shown that engineering microbial cocultures is an emerging and promising approach for biochemical production. In this study, we aim to explore the potential of ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-015-0319-0
更新日期:2015-09-15 00:00:00
abstract:BACKGROUND:Extracts and products (roots and/or aerial parts) from Echinacea ssp. represent a profitable market sector for herbal medicines thanks to different functional features. Alkamides and polyacetylenes, phenols like caffeic acid and its derivatives, polysaccharides and glycoproteins are the main bioactive compou...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-12-44
更新日期:2013-05-04 00:00:00
abstract:BACKGROUND:Under aerobic conditions, acetic acid is the major byproduct produced by E. coli during the fermentation. And acetic acid is detrimental to cell growth as it destroys transmembrane pH gradients. Hence, how to reduce the production of acetic acid and how to utilize it as a feedstock are of intriguing interest...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0475-x
更新日期:2016-05-05 00:00:00
abstract:BACKGROUND:Fatty aldehydes are industrially relevant compounds, which also represent a common metabolic intermediate in the microbial synthesis of various oleochemicals, including alkanes, fatty alcohols and wax esters. The key enzymes in biological fatty aldehyde production are the fatty acyl-CoA/ACP reductases (FARs)...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-018-0869-z
更新日期:2018-02-08 00:00:00
abstract:BACKGROUND:In spite of the substantial metabolic engineering effort previously devoted to the development of Saccharomyces cerevisiae strains capable of fermenting both the hexose and pentose sugars present in lignocellulose hydrolysates, the productivity of reported strains for conversion of the naturally most abundan...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-9-16
更新日期:2010-03-10 00:00:00
abstract:BACKGROUND:The budding yeast Pichia pastoris is widely used for protein production. To determine the best suitable strategy for strain improvement, especially for high secretion, quantitative data of intracellular fluxes of recombinant protein are very important. Especially the balance between intracellular protein for...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-10-47
更新日期:2011-06-26 00:00:00
abstract:BACKGROUND:Bacillus subtilis is an important industrial workhorse applied in the production of many different commercially relevant proteins, especially enzymes. Virtually all of these proteins are secreted via the general secretion (Sec) pathway. Studies from different laboratories have demonstrated essential or non-e...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-020-01315-2
更新日期:2020-02-28 00:00:00
abstract:BACKGROUND:The rational design of L-phenylalanine (L-Phe) overproducing microorganisms has been successfully achieved by combining different genetic strategies such as inactivation of the phosphoenolpyruvate: phosphotransferase transport system (PTS) and overexpression of key genes (DAHP synthase, transketolase and cho...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-6-30
更新日期:2007-09-19 00:00:00
abstract:BACKGROUND:The Gram-positive bacterium Bacillus subtilis has been widely used as a cell factory for the production of proteins due to its generally regarded as safe (GRAS) nature and secretion capability. Of the known secretory pathways in B. subtilis, the majority of proteins are exported from the cytoplasm by Sec pat...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0469-8
更新日期:2016-04-28 00:00:00
abstract:BACKGROUND:Crocin is a carotenoid-derived natural product found in the stigma of Crocus spp., which has great potential in medicine, food and cosmetics. In recent years, microbial production of crocin has drawn increasing attention, but there were no reports of successful implementation. Escherichia coli has been engin...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-019-1166-1
更新日期:2019-07-05 00:00:00
abstract:BACKGROUND:Cholesterol, the precursor of all steroid hormones, is the most abundant steroid in vertebrates and exhibits highly hydrophobic properties, rendering it a difficult substrate for aqueous microbial biotransformations. In the present study, we developed a Bacillus megaterium based whole-cell system that allows...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-015-0300-y
更新日期:2015-07-29 00:00:00
abstract:BACKGROUND:Bacillus subtilis has been widely used as a host for heterologous protein expression in food industry. B. subtilis ATCC6051 is an alternative expression host for the production of industrial enzymes, and exhibits favorable growth properties compared to B. subtilis 168. Extracellular expression of pullulanase...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-018-1011-y
更新日期:2018-10-22 00:00:00
abstract:BACKGROUND:The efficiency of biotechnological production processes depends on selecting the best performing microbial strain and the optimal cultivation conditions. Thus, many experiments have to be conducted, which conflicts with the demand to speed up drug development processes. Consequently, there is a great need fo...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-9-86
更新日期:2010-11-13 00:00:00
abstract:BACKGROUND:Gene therapy has gained an increasing interest in its anti-tumor efficiency. However, numerous efforts are required to promote them to clinics. In this study, a novel and efficient delivery platform based on bacterial magnetosomes (BMs) were developed, and the efficiency of BMs in delivering small interferin...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-017-0830-6
更新日期:2017-11-28 00:00:00
abstract:BACKGROUND:Trypsin has many applications in food and pharmaceutical manufacturing. Although commercial trypsin is usually extracted from porcine pancreas, this source carries the risks of infectivity and immunogenicity. Microbial Streptomyces griseus trypsin (SGT) is a prime alternative because it possesses efficient h...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-020-01338-9
更新日期:2020-04-13 00:00:00
abstract:BACKGROUND:Anaerobic digesters become unstable when operated at a high organi c loading rate (OLR). Investigating the microbial community response to OLR disturbance is helpful for achieving efficient and stable process operation. However, previous studies have only focused on community succession during different proc...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0466-y
更新日期:2016-04-25 00:00:00
abstract:BACKGROUND:The six-carbon circular non-proteinogenic compound L-pipecolic acid is an important chiral drug intermediate with many applications in the pharmaceutical industry. In the present study, we developed a metabolically engineered strain of Escherichia coli for the overproduction of L-pipecolic acid from glucose....
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-017-0666-0
更新日期:2017-03-27 00:00:00
abstract:BACKGROUND:L-tryptophan is an aromatic amino acid widely used in the food, chemical and pharmaceutical industries. In Escherichia coli, L-tryptophan is synthesized from phosphoenolpyruvate and erythrose 4-phosphate by enzymes in the shikimate pathway and L-tryptophan branch pathway, while L-serine and phosphoribosylpyr...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-11-30
更新日期:2012-03-02 00:00:00
abstract:BACKGROUND:Medium-chain-length polyhydroxyalkanoates (mcl-PHAs) containing various chain length monomers from C6 to C14 have more applications besides sustainable and environmental-friendly biomaterials owing to their superior physical and mechanical properties. We engineered a reversed fatty acid β-oxidation pathway i...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-019-1186-x
更新日期:2019-08-13 00:00:00
abstract:BACKGROUND:In our previous study, the feasibility of Rubisco-based engineered E. coli (that contains heterologous phosphoribulokinase (PrkA) and Rubisco) for in situ CO2 recycling during the fermentation of pentoses or hexoses was demonstrated. Nevertheless, it is perplexing to see that only roughly 70 % of the carbon ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0530-7
更新日期:2016-08-02 00:00:00
abstract:BACKGROUND:Membrane proteins are important drug targets in many human diseases and gathering structural information regarding these proteins encourages the pharmaceutical industry to develop new molecules using structure-based drug design studies. Specifically, membrane-bound catechol-O-methyltransferase (MBCOMT) is an...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-015-0304-7
更新日期:2015-08-07 00:00:00
abstract:BACKGROUND:E. coli is a robust host for various genetic manipulations and has been used commonly for bioconversion of hexose and pentose sugars into valuable products. One of the products that E. coli make under fermentative condition is ethanol. However, availability of limited reducing equivalence and generation of c...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-11-145
更新日期:2012-11-04 00:00:00
abstract:BACKGROUND:L-Alanyl-L-glutamine (AQ) is a functional dipeptide with high water solubility, good thermal stability and high bioavailability. It is widely used in clinical treatment, post-operative rehabilitation, sports health care and other fields. AQ is mainly produced via chemical synthesis which is complicated, time...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-020-01369-2
更新日期:2020-06-11 00:00:00
abstract:BACKGROUND:Purine nucleotides exhibit various functions in cellular metabolism. Besides serving as building blocks for nucleic acid synthesis, they participate in signaling pathways and energy metabolism. Further, IMP and GMP represent industrially relevant biotechnological products used as flavor enhancing additives i...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-11-138
更新日期:2012-10-24 00:00:00
abstract:BACKGROUND:Cost-effective production of the highly effective anti-cancer drug, paclitaxel (Taxol®), remains limited despite growing global demands. Low yields of the critical taxadiene precursor remains a key bottleneck in microbial production. In this study, the key challenge of poor taxadiene synthase (TASY) solubili...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-020-01458-2
更新日期:2020-11-02 00:00:00
abstract:BACKGROUND:Messenger RNA decay is an important mechanism for controlling gene expression in all organisms. The rate of the mRNA degradation directly affects the steady state concentration of mRNAs and therefore influences the protein synthesis. RNaseE has a key importance for the general mRNA decay in E. coli. While RN...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-5-38
更新日期:2006-12-12 00:00:00
abstract:BACKGROUND:Corynebacterium glutamicum was used as a metabolic engineering chassis for production of crude violacein (mixture of violacein and deoxyviolacein) due to Corynebacterium's GRAS status and advantages in tryptophan fermentation. The violacein is a commercially potential compound with various applications deriv...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0545-0
更新日期:2016-08-24 00:00:00
abstract:BACKGROUND:Protoilludene is a valuable sesquiterpene and serves as a precursor for several medicinal compounds and antimicrobial chemicals. It can be synthesized by heterologous expression of protoilludene synthase in Escherichia coli with overexpression of mevalonate (MVA) or methylerythritol-phosphate (MEP) pathway, ...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0409-7
更新日期:2016-01-19 00:00:00