Supplementary Table 1: Surface analysis by subfamily annotation of organic solvent stable lipases

Subfamily 1 / Temperature stability / pH stability / Organic stability / Substrate / Application / LogP / *Surface properties
SNL / Mesostable / Alkalophilic / Petroleum spirit, heptane, toluene, DMF, and DMSO, di-isopropylfluorophosphate.
Unstable in high concentration of PMSF, SDS, acetone, benzene, ethanol, and methanol
EDTA has no affect. / Long chain / Organic synthesis / Wide / N and Ne residues more SE. Surface exposure of D > E
Res less Hp till 70-80%. From 60-70% till 30-40% more P and H SE.
No C present. W and H buried.
Q> SE. A more buried. L more buried.
Subfamily 2 / Temperature stability / pH stability / Organic stability / Substrate / Application / LogP / *Surface properties
YLL / Thermostable / 8 / Water miscible organic solvents, 10% acetone, methanol , ethanol, isopropanol, DMSO. Maximum activity in DMSO.
In 20% organic solvents, there was no activity left in acetone, ethanol, isopropanol, 60% activity remaining in methanol and 95% activity remaining in DMSO. / Long chain
C12-c16 / Biodissel, organic synthesis / -1.12-0.9 / More Ne and N SE. Hp less SE. C29 20-30%, C42 20-30% and C243 10-20% SE. 2 C buried. M100 50-60% SE. K and H more SE exposed than R. More D SE. W SE. G more buried. Q>SE. 2 DSB between residues 42-298 and 119-112.
L more buried.
ANLO44 / Thermostable / Acidic-neutral / Stable in Methanol, 2-propanol & methyl acetate
No deactivation in presence of 50 % water-miscible organic solvents. However, water-immiscible
aliphatic solvents reduce the lipase activity between 20 and 80 %. / 1,3 regiospecific
Middle chain / Biodissel, organic synthesis / -0.55 -0.25 / Ne and N SE, Hp and P less SE. C104 30-40%, C35 10-20% SE. 5 cys buried. D>E SE. > K and H SE than R.. M209 30-40% SE. M132 & M50 burtied. W SE.
ATL / Wide range of stability / 3-12
stable over 4-10 / Stable in various organic solvents
Inhibited by ionic detergents, whereas non-ionic detergents stimulates enzyme activity.
Unaffected by EDTA or by 2-mercaptoethanol and potassium ferrocyanide / 1,3 regiospecific / Wide / Complicated exposure of surface charges. C273 and C110 20-30% and 109-20% SE respectively. 4 cys buried. D and E equally SE. K res SE. M1 60-70% SE. H< SE. W SE.
1 DSB between residues 110-113.
FHL / Thermostable / Acidic-neutral / 50% DMSO, hexane, benzene. / Middle chain fatty acid
1,3- regiospecific / Organic synthesis / -1.12- 3.5 / More Ne SE till 60-70%. N and Hp balance each other. D SE. > K SE. H buried. M243 60-70% SE. M300 40-50% SE. M292 20-30% SE. 1M buried. 6 C buried. W SE. 1 DSB between residues 36-41. A, L buried
ROL / Mesostable / Neutral / Fairly stable in alkanes and long chain alcohols. Most stable in hexane.
Denatured in hydrophilic solvents such as acetone or short chain (C1–C3) alcohol. Most stable in hexane.
CTAB reduces enzyme activity / Medium chain fatty acids; hydrophobic, crevice-like binding site located near the protein surface flexible substrates are preferably
hydrolyzed in sn-1, rigid substrates in sn-3
Do not bind tertiary alcohol / Organic synthesis / Log P >2 / P and N more SE. C266 30-40% SE. C38 20-30% SE. M154 buried. No SE M. H res buried. More K SE. >D SE. 4 C buried. W more SE. Maximum Q SE. 1 DSB between residues 27-38.
RML / Mesostable / Alkalophilic / 80 mM Tween 80 or 90 mM Tween 20, hexane. / -do-
Bind tertiary alcohol / Detergent
Organic synthesis / >4 / N and Hp more SE. Ne more SE. D>E SE. His SE. W SE. Y, M buried. 1 DSB between residues 182-240.
Subfamily 3 / Temperature stability / pH stability / Organic stability / Substrate / Application / LogP / *Surface properties
PFL / Thermostable / Alkalophilic / Water-miscible organic solvents solutions and highly tolerant to some water-immiscible organic solvents.
Stable in isopropanol, methanol and ethanol, as well as acetone and glycerol
Inactivated in cyclohexane, chloroform, benzene and acetic acid. / - / Many / Unable to predict / R res more SE. C buried. E>D SE. P and N more SE. W SE. His SE. Q more buried than SE. Q more buried.
ASYL / Thermostable / Alkalophilic / Most active in methanol and have 60% of this activity in hexane. / - / Biodissel / -0.55-3.5 / Ne and Hp more SE. P and N more buried than SE. C15 and C20, 30-40% SE, C214 and C264 SE. E>D SE. K. M is absent. W buried. H buried. 1 DSB between residues 214-264.
PAL / Mesostable / - / Stable in n-hexane cyclohexan, p-xylene,
1-octanol, ethanol, pentanol inhibit it. / - / Organic synthesis / >2 / C183 and C235 form disulphide bond and are 30-40% SE. Ne more buried. N, P and Hp more SE. Two M 20-30% SE. most and more SE are R res. E>D SE. H more buried. W buried. His buried. 1 DSB between residues 183-235.
PLKW156 / Thermostable / Neutral / Stable in >95% Acetone, methanol and ethanol
Inhibited by anionic surfactants such
as SDS and LBS (sodium laurylbenzenesulfonate). / Medium and the long-acyl chain esters funnel-like substrate-binding sites / Biodissel / >0 / P and Ne less SE. N and Hp more SE. H buried. C194 and C274 40-50 and 30-40% SE respectively. 1 M present which is buried. H158 90-100 SE. W buried. More H buried. Q< SE.
PCL / thermostable / Neutral / Stable in methanol, ethanol, acetone, acetonitrile, DMF, DMSO and dioxane
Benzene or n-hexane had inhibitory effect / Short and middle chain fatty acids (<C 12) funnel-like substrate-binding sites / Organic synthesis
Biodissel / -1.12-0.17 / 2 C surface exposed C190 and C270 which forms DSB. N and H more SE. Ne more shielded than P. M255 Buried. H and D res are buried. E>D SE. However D>E. W buried. All H buried. Q< SE. 1 DSB between residues 190-270.
RLM / Thermostable / Alkalophilic / Stable in tert-butanol, hexane dichloromethane, ethyl acetate, ethanolamine
In DMSO and methanol showed no activity.
Acetonitrile and 1-butanol inhibitory
DMFA, acetone, ethanol, and isopropanol decreased activity / - / Organic synthesis / Unable to predict / N and Hp more SE. D>E SE. N more SE. C240 20-30% SE.
C182 40-50% SE. All H and M buried. Ne more SE than P at 60-70% & 10-20%. P more in 10-30%.
More L SE. W buried. Q SE
1 DSB between residues 182-240.
PS5L / Thermostable / Alkalophilic / n-hexane, tween -60, tween-80 / - / Organic synthesis, detergent / >2 / Max C SE. H res very less those present are SE. R>K SE. E res more SE. Out of 12 C 8 are SE. P>Ne Hp and N are equivalently distributed. Maximum Y buried. W SE. F more buried. H SE.
Subfamily 4 / Temperature stability / pH stability / Organic stability / Substrate / Application / Log P / *Surface properties
GthLYN / Thermostable / Alkalophilic / Stable in 1%sds edta and mercaptoethanol, ethanol.
DTT, DMSO and isopropanol showed slight inhibition / Medium chain length triglycerides / Biodissel, organic synthesis / Unable to predict / Frequency of Hp and N SE. P> Ne SE. H140 and H312 50-60% SE. M351 10-20% SE. H153 10-20% SE. H85 and H152 20-30% SE. 5 M and 2 C res buried. R> K SE. H res equivalently SE and buried. W SE.Y more SE. L more buried.
BTL / do / do / Cholate, CHAPS, ethanol for 1 hour at 30 degree then decreases / Medium chain length triglycerides / Detergent, environment / >2<4 / C296 and C65 buried. M174 10-20% SE. 6 His SE, 7 buried. 4 M buried. > number of D SE but E more SE than D. P more SE. Hp and N equal contribution at 70-90 and 40-50%, At 20-30% >Hp res. W SE. Y & F> SE.
BL42 / do / do / Stable benzene hexane, decanol, isooctane, tetradecane water immiscible solvents enhanced activity.
Polar solvents reduced activity / Medium chain length triglycerides
low phospholipase activity / Detergent, organic synthesis, biodissel / 2
More stable in >4 / P > SE. > N and Hp than P and Ne. M173 10-20% SE. 5 M and 2 C buried. 6 H SE and 6 buried. W shows equivalent distribution. Y > SE.
BSLP1 / do / do / Chaps and Triton X-100
SDS, Tween20 inhibitted / Short-Medium chain length triglycerides / Detergent, environment / <4 / N res moreSE. P more Se. More D SE. R >K in SE. H312 60-70% SE, H140 and H223 50-60% SE. H153 40-50% SE, H85 and H152 20-30% SE. 2 M 10-20% SE. 2 C buried. 8 H and 3 M buried. W buried. >F buried. >Y SE. > L buried.
GLTW1 / do / do / DTT, SDS, 1%detergents (Tween 20, Chaps or Triton X-100) / Medium chain length triglycerides / Dertergent, Environment / >2 / Maximum N SE. P> SE. Ne more in 70-80%, 60-70% and 10-20%. M173 10-20% SE. H223, H140, H152, 153, 85, 87 SE. 5 H res buried, C64 and C295 buried. 5 M buried. D>E SE. > W SE. > L buried. > M buried.
SSL / - / - / p-xylene, benzene, toluene and hexane / - / Organic synthesis / >2 / C98 80-90% SE. C22 40-50% SE. C25 30-40% SE. C343 10-20% SE. 3 C buried. 2 M buried. 4 H SE and 8 H buried. D more SE than E. K more SE than R. N>Hp SE. W > buried. Y> SE.
SAL3 / Thermostable / Alkalophilic / 20mM Triton and TX-100, NaDC, NaTDC, and Tween 80, H2O2, Sodium
perborate. / Short-chain fatty acid esters / Organic synthesis, detergent, environment / >2 / P Ne SE. N more SE. K > R. H274, H146, H300, H143, H158, H55 are 70-80, 60-70, 50-60, 30-40 and 10-20% SE. M256 50-60%, M104, M256 10-20% SE. 6 M buried. E more SE. > Y SE. W SE. > G SE.
Subfamily 5 / Temperature stability / pH stability / Organic stability / Substrate / Application / Log P / *Surface properties
BSL / Mesostable / Alkalophilic / 1-butanol, hexanol, benzene and toluene / Short chain triglycerides / Biodissel
Organic synthesis / Around 2 / P more SE. N res more SE. C absent. Hp more in 50-60%. M134, M137 and M78 SE. H152, H3, H156 SE. 1 M and 2 H. res buried. K>SE. D more and > SE. W SE. Y> SE.
BLL / Thermostable / Alkalophilic / Various detergents and organic solvents / Short chain triglycerides / Varied / -  / N res > SE. P res more in 60-70% and 10-30%. No C present. 3 M SE. 2 M buried. >K SE. R more SE than K. H184, H35 H188 SE. 2 H res buried. W SE
Subfamily 6 / Temperature stability / pH stability / Organic stability / Subtrate / Application / LogP / *Surface properties
PLB-11 / Mesostable / Alkalophilic / Ethanol, Methanol, Dimethyl sulphoxide, Dimethyl formamide / Short chain triglycerides / Detergent, biodissel / -1.12- -0.33 / 3 C and 1 M buried. C136 30-40% SE. C79 20-30% SE. C221 10-20% SE. Ne res more SE than P. N res more SE. E > D SE. >W SE. >H SE. >L buried.> M SE.
BSL205y / Moderate thermostable / Neutral / p-xylene, n-decane
Complete deactivation in acetonitrile
No activity in hexadecane / Short chain triglycerides / Organic synthesis / >2-4 <4 / H206 50-60 SE. H104 30-40% SE. H 340 and M81 10-20% SE. C absent. 6 M buried. 3 H res buried. N res more SE. Hp less SE. Nothing conclusive can be said about P and Ne residues. > W buried. 7 L SE, 11 buried. >F SE. >M buried.
Subfamily 7 / Temperature stability / pH stability / Organic stability / Substrate / Application / LogP / *Surface properties
TTL / Thermostable / Alkalophilic / tert-butanol, acetonitrile, isopropanol, pyridine, acetone up to a concentration of 99% (v/v) , DTT (dithiotreitol), guanidine hydrochloride, pHMB (phydroxymercuribenzoate) and iodo-acetate / Medium chain length triglycerides / Varied / -0.34- 0.77 / No C. Ne> P SE. K more SE and > R. H19 10-20%, H233 20-30%, H169 50-60%, H45 70-80%, SE. 6 M buried. M147 60-70%, M1 50-60%, M158 20-30%, M114 10-20 , M17 SE. W more buried. L<SE. H> SE. Y>SE absent in core. G>SE. Q very less and least SE.
Subfamily 8 / Temperature stability / pH stability / Organic stability / Substrate / Application / Log P / *Surface properties
SRL / Thermostable / Alkalophilic / Water miscible organic solvents, acetone, methanol, ethanol, isopropanol, DMSO / - / Biodissel, detergent, organic synthesis / >2 / Ne > SE. Hp < SE. Ne>P. C288 20-30% SE. C276 10-20% SE. M518 20-30% SE. D>E SE. M523 40-50% SE. H370, H491 40-50%H298, H453, H243, H41, H492 20-30%, H188 10-20% SE. 5 M and 2 C buried. R are buried but K SE. W<SE. F<SE. L, A, V more buried.
Subfamily 9 / Temperature stability / pH stability / Organic stability / Substrate / Application / Log P / *Surface properties
GGYO5l / Moderate thermostable / Neutral-alkaline / Methanol, glycerol, hexane,n-heptane, cyclohexane, isooctane, benzene, and diethyl ether, DMSO / - / Biodissel, detergent, organic synthesis / >2 / Ne > SE. Hp < SE. Ne>P. C288 20-30% SE. C276 10-20% SE. M518 20-30% SE. D>E SE. M523 40-50% SE. H370, H491 40-50%H298, H453, H243, H41, H492 20-30%, H188 10-20% SE. 5 M and 2 C buried. R are buried but K SE. W<SE. F<SE. L, A, V more buried.
Subfamily 10 / Temperature stability / pH stability / Organic stability / Substrate / Application / Log P / *Surface properties
SML / Moderate thermostable / Neutral-alkaline / Various water miscible and immiscible / - / Varied / <2 / N SE. NeSE than P. Hp res less SE. KSE. M1 30-40% and M324 10-20% SE. 1 M buried. C absent D SE than E. W maximum SE. F<SE.

Hp-hydrophobic, N- neutral, P-positive, Ne-negative residues, DSB- Disulphide bond, SE- Surface exposed and res-residues. Amino acid is represented by their single letter codes * Surface properties were analyzed through ASAview server. – represents unknown.

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