Bacterial Hormone-Sensitive Lipases (bHSLs): Emerging Enzymes for Biotechnological Applications

( T. Doohun Kim ) 한국미생물생명공학회(구 한국산업미생물학회) 2017 Journal of microbiology and biotechnology Vol.27 No.11

Lipases are important enzymes with biotechnological applications in dairy, detergent, food, fine chemicals, and pharmaceutical industries. Specifically, hormone-sensitive lipase (HSL) is an intracellular lipase that can be stimulated by several hormones, such as catecholamine, glucagon, and adrenocorticotropic hormone. Bacterial hormone-sensitive lipases (bHSLs), which are homologous to the C-terminal domain of HSL, have α/β-hydrolase fold with a catalytic triad composed of His, Asp, and Ser. These bHSLs could be used for a wide variety of industrial applications because of their high activity, broad substrate specificity, and remarkable stability. In this review, the relationships among HSLs, the microbiological origins, the crystal structures, and the biotechnological properties of bHSLs are summarized.

Structural and functional analyses of a bacterial homologue of hormone-sensitive lipase from a metagenomic library.

Ngo, Tri Duc,Ryu, Bum Han,Ju, Hansol,Jang, Eunjin,Park, Kwangsoo,Kim, Kyeong Kyu,Kim, T Doohun Wiley-Blackwell 2013 Acta crystallographica. Section D, Biological crys Vol.69 No.9

<P>Intracellular mobilization of fatty acids from triacylglycerols in mammalian adipose tissues proceeds through a series of lipolytic reactions. Among the enzymes involved, hormone-sensitive lipase (HSL) is noteworthy for its central role in energy homeostasis and the pathogenic role played by its dysregulation. By virtue of its broad substrate specificity, HSL may also serve as an industrial biocatalyst. In a previous report, Est25, a bacterial homologue of HSL, was identified from a metagenomic library by functional screening. Here, the crystal structure of Est25 is reported at 1.49?? resolution; it exhibits an α/β-hydrolase fold consisting of a central β-sheet enclosed by α-helices on both sides. The structural features of the cap domain, the substrate-binding pocket and the dimeric interface of Est25, together with biochemical and biophysical studies including native PAGE, mass spectrometry, dynamic light scattering, gel filtration and enzyme assays, could provide a basis for understanding the properties and regulation of hormone-sensitive lipase (HSL). The increased stability of cross-linked Est25 aggregates (CLEA-Est25) and their potential for extensive reuse support the application of this preparation as a biocatalyst in biotransformation processes.</P>

Lipid-lowering effects of Zanthoxylum schinifolium Siebold & Zucc. seed oil (ZSO) in hyperlipidemic rats and lipolytic effects in 3T3-L1 adipocytes

Kim, Kyoung Kon,Kim, Tae Woo,Kang, Yun Hwan,Kim, Dae Jung,Choe, Myeon Korean Society of Food Science and Technology 2016 Food Science and Biotechnology Vol.25 No.5

On the basis of the antiatherosclerotic effect of Zanthoxylum schinifolium, the therapeutic potential of Zanthoxylum schinifolium seed oil (ZSO) was tested in terms of the blood lipid profile and obesity in rats. The lipolytic effects of ZSO were determined in adipocytes and the total body and liver weight were decreased in rats. Compared with the high-cholesterol high-fat (HCHF) group, the rats in the HCHF+ZSO group showed improved levels of hyperlipidemia indicators. Furthermore, western blot analysis confirmed that the improvement of hyperlipidemia indicators was induced by stimulation of lipoprotein lipase expression. Additional results indicated that the reduction in body weight was likely caused by phosphorylation of hormone-sensitive lipase (HSL) via the protein kinase A pathway, ultimately leading to lipolysis. In conclusion, the results of the in vivo experiment showed that ZSO improved the lipid profiles in the blood, lowering cardiovascular disease and arteriosclerosis and degrading cellular lipids by activating HSL.

Structural and functional analysis of a novel hormone-sensitive lipase from a metagenome library

Nam, Ki Hyun,Kim, Min-Young,Kim, Soo-Jin,Priyadarshi, Amit,Kwon, Suk-Tae,Koo, Bon-Sung,Yoon, Sang-Hong,Hwang, Kwang Yeon Wiley Subscription Services, Inc., A Wiley Company 2009 Proteins Vol.74 No.4

Lipid-lowering effects of Zanthoxylum schinifolium Siebold & Zucc. seed oil (ZSO) in hyperlipidemic rats and lipolytic effects in 3T3-L1 adipocytes

김경곤,김태우,강윤환,김대중,최면 한국식품과학회 2016 Food Science and Biotechnology Vol.25 No.5

On the basis of the antiatherosclerotic effect of Zanthoxylum schinifolium, the therapeutic potential of Zanthoxylum schinifolium seed oil (ZSO) was tested in terms of the blood lipid profile and obesity in rats. The lipolytic effects of ZSO were determined in adipocytes and the total body and liver weight were decreased in rats. Compared with the high-cholesterol high-fat (HCHF) group, the rats in the HCHF+ZSO group showed improved levels of hyperlipidemia indicators. Furthermore, western blot analysis confirmed that the improvement of hyperlipidemia indicators was induced by stimulation of lipoprotein lipase expression. Additional results indicated that the reduction in body weight was likely caused by phosphorylation of hormone-sensitive lipase (HSL) via the protein kinase A pathway, ultimately leading to lipolysis. In conclusion, the results of the in vivo experiment showed that ZSO improved the lipid profiles in the blood, lowering cardiovascular disease and arteriosclerosis and degrading cellular lipids by activating HSL.

A New Esterase, Belonging to Hormone-Sensitive Lipase Family, Cloned from Rheinheimera sp. Isolated from Industrial Effluent

Virk, Antar Puneet,Sharma, Prince,Capalash, Neena The Korean Society for Microbiology and Biotechnol 2011 Journal of microbiology and biotechnology Vol.21 No.7

The gene for esterase (rEst1) was isolated from a new species of genus Rheinheimera by functional screening of E. coli cells transformed with the pSMART/HaeIII genomic library. E. coli cells harboring the esterase gene insert could grow and produce clear halo zones on tributyrin agar. The rEst1 ORF consisted of 1,029 bp, corresponding to 342 amino acid residues with a molecular mass of 37 kDa. The signal P program 3.0 revealed the presence of a signal peptide of 25 amino acids. Esterase activity, however, was associated with a homotrimeric form of molecular mass 95 kDa and not with the monomeric form. The deduced amino acid sequence showed only 54% sequence identity with the closest lipase from Cellvibrio japonicus strain Ueda 107. Conserved domain search and multiple sequence alignment revealed the presence of an esterase/ lipase conserved domain consisting of a GXSXG motif, HGGG motif (oxyanion hole) and HGF motif, typical of the class IV hormone sensitive lipase family. On the basis of the sequence comparison with known esterases/ lipases, REst1 represents a new esterase belonging to the class IV family. The purified enzyme worked optimally at $50^{\circ}C$ and pH 8, utilized pNP esters of short chain lengths, and showed best catalytic activity with p-nitrophenyl butyrate ($C_4$), indicating that it was an esterase. The enzyme was completely inhibited by PMSF and DEPC and showed moderate organotolerance.

Effects of Dietary Energy Density on Growth, Carcass Quality and mRNA Expression of Fatty Acid Synthase and Hormone-sensitive Lipase in Finishing Pigs

Liu, Z.H.,Yang, F.Y.,Kong, L.J.,Lai, C.H.,Piao, X.S.,Gu, Y.H.,Ou, X.Q. Asian Australasian Association of Animal Productio 2007 Animal Bioscience Vol.20 No.10

A single factorial experiment was conducted to test the effects of three dietary levels of energy on mRNA expression of fatty acid synthase (FAS-mRNA) and hormone-sensitive lipase (HSL-mRNA) and their association with intramuscular fat in finishing pigs. 72 crossbred (Large $White{\times}Rongchang$) barrows with an average initial body weight of 20.71 (s.e. 0.1) kg, were randomly allotted to three dietary treatments (11.75, 13.05 and 14.36 MJ DE/kg) and fed until slaughtered at 100 or 101 kg. The diets were iso-nitrogenous and iso-essential amino acids. The growth performances including the duration of finishing were changed linearly (p<0.05) or quadratically (p<0.05) with increased dietary energy levels. The effects of dietary energy content on the percentage of external fat, intramuscular backfat and the fat thickness were linear (p<0.05). The content of dietary energy increased FAS-mRNA linearly or quadratically, while HSL-mRNA decreased linearly or quadratically in backfat and Longissmus dorsi muscle. Meanwhile, significant positive correlations (p<0.05) were found between energy level and intramuscular fat, FAS-mRNA or the ratio of FAS-mRNA to HSL-mRNA, between the ratio of FAS-mRNA to HSL-mRNA and intramuscular fat. However, the correlations between HSL mRNA and dietary energy or intramuscular fat were negative (p<0.05). The results indicated that dietary energy level regulates lipid accumulation, especially intramuscular fat, possibly by modulating the mRNA of FAS and HSL together rather than individually.

유색미 겨 아라비녹실레인 섭취와 지구성 운동트레이닝이 흰쥐의 근섬유 유형별 HSL과 LPL 단백질 발현에 미치는 영향

여효성(Yeo, Hyo-Sung),이상호(Lee, Sang-Ho),손희정(Son, Hee-Jeong) 한국체육과학회 2015 한국체육과학회지 Vol.24 No.5

Arabinoxylan is one of the potential key products of a rice bran based biorefinery. Arabinoxylan rice bran has been shown to be a potent biological response modifier. In most country, obesity and related diseases have emerged as major health problems. Hormone-sensitive lipase (HSL) is important for the degradation of triacylglycerol in adipose and muscle tissue, but the tissue-specific regulation of this enzyme is not fully understood. We investigated the effects of Ara rice bran and exercise on the fat metabolism in rats. As the results, HSL as a major enzyme involved in triacylglycerol lipolysis showed a significantly higher in exercise group and Ara supplement group and Ara supplement with exercise group in EDL muscle. Whereas, LPL(lipoprotein lipase) as a second enzyme in lipolysis showed a significantly lower in Ara supplement with exercise group, thus the Ara supplement with exercise group had a higher inhibitory activity than the both exercise and Ara supplement group in EDL muscle. However, HSL enzyme in all group significantly decreased than control group in soleus muscle. These results require further research and suggest that the supplement of Ara rice bran with exercise is likely contribute to triacylglycerol lipolysis response in only EDL muscle of rat. Through a further study, the effect of ara rice bran can be confirmed as a possible safe alternative to the treatment modalities.

Aronia melanocarpa reduced adiposity via enhanced lipolysis in high-fat diet-induced obese mice

Kim, Hyun Kyung,Jung, Jiyeon,Kang, Eun Young,Gang, Gyoungok,Kim, Wooki,Go, Gwang-woong Korean Society of Food Science and Technology 2020 한국식품과학회지 Vol.52 No.3

Obesity is a critical health issue in Korea, where half of all adults are overweight and a third obese. Aronia melanocarpa -rich in flavonoids and phenolics- with antioxidant and anti-inflammatory activities, could have anti-obesity activity and reduce body fat mass by upregulating lipolysis and β-oxidation in obese mice. Male C57BL/6J mice (n=12) were assigned into four groups: normal chow (18% kcal from fat); high-fat diet control (HFD, 45% kcal from fat); HFD+A. melanocarpa (200 mg/kg diet); HFD+Xenical (500 mg/kg diet, positive control). Antioxidant capacity of A. melanocarpa was established in vitro and in vivo. Weight loss was induced as decreased adiposity and lowered respiratory quotient at rest suggested oxidation of stored fat. Adiposity reduction, accompanied with elevated fat utilization, was owing to enhanced activity of hormone-sensitive lipase. Thus, A. melanocarpa lowered adiposity by enhancing lipolysis and utilization of fatty acids in visceral fat.

Anti-adipogenic Effect of Undaria pinnatifida Extracts by Ethanol in 3T3-L1 Adipocytes

HyeJin Kim(김혜진),Chang-Han Kang(강창한),Sung-Koo Kim(김성구) 한국생명과학회 2012 생명과학회지 Vol.22 No.8

미역(Undaria pinnatifada)은 낮은 칼로리 및 요오드의 원료로써 천연체중조절식품으로 알려져 있다. 미역이 체중조절식품으로 알려져 있음에도 불구하고, 지방세포 분화 및 지방축적에 관한 저해 기작은 연구가 미비하다. 본 연구에서는 3T3-L1에서 지방세포로 분화가 일어나는 단계에서 미역에탄올추출물의 효과 및 기작을 확인하였다. 미역에탄올추출물의 독성과 지방축적저해효과는 MTT assay, Oil red O staining, RT-PCR과 western blot으로 분석하였다. 미역에탄올추출물은 50 μg/ml의 농도에서 독성을 띄지 않았다. 3T3-L1의 분화 및 지방세포에서 triglyceride축적과정동안 50 μg/ml의 미역에탄올추출물을 처리하였으며, 미역에탄올추출물은 지방세포에서triglyceride의 축적을 40% 감소시켰다. 지방세포 특이적 단백질인 Peroxisome proliferator activated receptor γ (PPARγ), leptin과 Hormone sensitive lipase (HSL)의 발현은 RT-PCR과 western blot으로 확인하였다. PPARγ의 과발현은 지방세포의 분화를 촉진시킨다. 또한 지방세포 크기의 증가와 세포 내 triglyceride의 함량에 따라 leptin은 세포 외로 분비된다. 그러므로 PPAR γ와 leptin은 비만의 지표로 사용된다. 첨가한 미역에탄올추출물의 농도가 높아질수록 PPARγ와 leptin의 발현이 억제되었다. 이상의 결과를 통하여, 미역의 에탄올 추출물은 지방전구세포의 분화를 억제시키며, 지방세포 내 triglyceride의 축적을 저해하는 것으로 판단된다. Undaria pinnatifada has been used as a natural diet food with few calories and as a source of iodine. Even though U. pinnatifida has been regarded as a diet food, the mechanisms of its inhibitory effects on adipocyte differentiation and the accumulation of fat in adipocytes are poorly understood. In this study, the effect and mechanism of U. pinnatifida ethanol extract on 3T3-L1 differentiation into adipocytes were investigated. The effects of U. pinnatifida ethanol extract on cell viability and the anti-adipogenic effect were investigated via MTT assay, Oil red O staining, RT-PCR, and western blot. The U. pinnatifida ethanol extract did not show toxicity up to a concentration of 50 μg/ml. The addition of U. pinnatifida ethanol extract decreased triglyceride contents by 40% when 50 μg/ml of U. pinnatifida ethanol extract was added during 3T3-L1 differentiation and adipocyte triglyceride formation. The transcription and expression of peroxisome proliferator-activated receptor γ (PPARγ), leptin, and hormone-sensitive lipase (HSL) as adipocyte-specific proteins were determined by RT-PCR and western blot. The overexpression of PPARγ could accelerate adipocyte differentiation. Also, leptin was secreted for triglyceride accumulation in the adipocytes and the increase of adipocyte cell size. Thus, PPARγ and leptin were used as indicators of obesity. PPARγ and leptin were repressed by the increased addition of U. pinnatifida ethanol extract. This indicates that U. pinnatifida was effective as an anti-obesity agent by repressing the differentiation of 3T3-L1 into adipocytes and inhibiting triglyceride formation in adipocytes.