yyareriyeaaiis Headtosavemyexams.co ukfor more awesome resources 2.10 Role of Enzymes Enzymes as Biological Catalysts Enzymes + Enzymes areproteins thatactas biological catalysts to speed up the rate of a chemical reaction without being changed or used up in the reaction * They are biological because they are madein living cells * Enzymes arenecessary toall living organisms as they maintain reaction speeds of all metabolic reactions atarate thatcan sustain life © Forexample, ifwedidnot produce digestive enzymes, itwouldtake around2-3weeks to digest one meal; with enzymes, ittakes around 4 hours © Often the products of one reaction arethe reactants foranother(andso on) The mechanism of enzyme action . + Enzymesarespecific to one particularsubstrate(s) as the active site of the enzyme, where the substrate attaches, is a complementary shape to thesubstrate + When the substrate moves intothe enzyme's activesite they become known as the ‘enzyme-substrate complex * After the reactionhas occur'ed, the products leave the enzyme’s active siteas they no longer fititanditis free to take upanothersubstrate © Step One: Enzymes and substrates randomly move about in solution © Step Two: When anenzyme andits complementary substrate randomly collide an enzyme-substrate complex forms, and the reaction occurs, © Step Three: A product (or products} forms from the substate(s) which are then released from theactve site. The enzymeis unchanged and wil goonto catalyse funherreactions w STARE specie. T ENZYME 4 ( .) ENZYME 4 IS COMPLEMENTARY IN SHAPE TO THE ‘SUBSTRATE ENZYME 2 IS NOT ABLE TO BIND TO THIS SUBSTRATE:Prey AME NUL COMPLEMENTARY. Ba SUBSTRATE AND ENZYME JOIN LIKE A ‘LOCK AND KEY’ PRODUCTS ENZYME IS UNCHANGED: AT THE END OF THE REACTION a sete aie} Factors Affecting Enzyme Action: Temperature + Enzymes are proteins andhavea specific shape, determined by the amino acids that make the enzyme and heldin place by bonds * This is extremely important around the active sites the specific shapeis what ensures the substrate will ftinto the active site and enable the reaction to proceed + Enzymes work fastest at their‘ optimum temperature’ © Inthehuman body, the optimum temperatureis 37°C + Heating to high temperatures (beyond the optimum) willbreak the bonds that hold the enzyme together anditwillloseits shape © This is known as denaturation + Substrates cannot fit into denatured enzymes as the shape of theiractive sitehas been lost + Denaturation is irreversible - once enzymes are denatured they cannot regain their proper shape andactivity willstopWe ACTIVE SITE 1 FORCES BETWEEN AMINO ACIDS IN THE CHAIN HOLD THE ENZYME TOGETHER AND CREATE ITS UNIQUE SHAPE AMINO ACID CHAN MAKING UP ENZYNE THE SUBSTRATE NO LONGER FITS THE SHAPE OF THE ACTIVE SITE HAS BEEN DISTORTED HGH TEMPERATURES BREAK THE FORCES THAT HOLD THE ENZYME TOGETHER Pasavemyexarns Effect of temperature on enzyme activity + Increasing the temperature towards the optimumincreases the activity of enzymes asthe morekineticenergy the molecules have the faster they move and thenumber of collisions with the substrate molecules increases, leading to a fasterrate of reaction AZyMes, they just make them work + This means that low temperatures do not denatur moreslowly due to alack ofkineticenergy& i Psavemyoxoms Graph showing the effect of temperature on the rate of enzymeactivity Practical: Enzymes & Temperature Amylaseis an enzyme that digests starch (a polysaccharide of glucose)intomaltose(a disaccharide of gucose) The effect of emperatureon the activityof amylase canbe investigated Apparatus Spotting ile Measuring cylinder Testtube Syringe Pipette Stopwatch Water Thermometer Waterbath lodine Starch solution ‘AmylasesolutionMethod + Add Sem? starch solution toa test tube andheat toa set temperatureusing beaker of \waterwthaBunsenoumer + Adda dropof lodineto¢ach of thewells of a spotting te + Use syringe to add 2crn* amylase to the starch solution and mixwell + Everyminute,transtera droplet of solution toa new well of iodine solution (which should tuin blue-black) + Repeat this transferprocess unti the iodine solution stops turning blue-black (this means the amylasehas broken downallthestarch) + Record the time taken for the reaction to be completed + Ropoat theinvestigation fora range of temperatures (from 20°C 10 60°C) THERMOMETER: a (SPOTTING: TeSee 4 é STARCH SOLUTION ‘ar sot ° 3 SSo8| [steers e eoee mae | [aot] [Ree = 9 MINUTES ‘3 MINUTES ELSOLDIGESTIONResults and Analysis + Amylase is an enzyme which breaks down starch + The quickerthereactionis completed, the faster the enzymes working + This investigation shows: © Atthe optimum temperature, the iodine stopped tumingblue-black the fastest + This is because the enzymeis working atits fastest ate anchas cigestedallthe starch © Atcolder temperatures (below optimum), teiodinetooks longertimeto stopturning blue-black * Thisis because the amylase enzymes working slowly due to lowkinetic energy and few collisions between the amylase andthe starch © Athotter temperatures (above optimum) the cine tumed blue-black throughout the wholeinvestigation © This is because the amylase enzyme has become denaturedand socanno longer bine with the starch orbreakit down Limitations + Themethodusedio controlthe temperature, described above, is not vary precise, an improvement wouldbe tousewater baths kept at each temperature" + The starch and amylase solutions that need tobe used shouldbe placed ina water’sath andallowedto each the temperature (using s thermometer to check) before beingused + Acolorimeter can boused tomeasure the progress of thereaction more accurately © solution containing starch willbe darker than solution contahing glucose(as resultof thecolourchange ofiocine) © Theabsorbance or transmission of ight through the coloured solution can be measuredusinga colorimeter Applying CORMS to practical work + When working with practicalinvestigations, remember toconsider your CORMS evaluationWHAT 1S BEING CHANGED IN THE INVESTIGATION (l.V) ‘A CONTROL RELATED TO THE ORGANISM BEING USED (IF RELEVANT) REPEATS MUST BE CARRIED OUT FOR RELIABLE RESULTS HOW WILL YOU MEASURE YOUR DEPENDENT] VARIABLE (D.V.) WHAT TIME SCALE WILL YOU USE (D.V.) WHAT WILL YOU CONTROL IN THE INVESTIGATION (C.V.1 CORMSevaluation + In thisinvastigation, your evaluation should iook something like this: © C-Wearechangingthe temperaturein each repeat, (O- Thisisnot relevant to this investigation as wearen’tusingan organism R_Wewilrepeat the investigationseveraltimes tomake sure ourresultsarereliable M1-Wewilmessurethetimetaken | * M2 forthe iodine to stop turning black 'S-Wewillcontrolthe concentration and volume of starch solution, iodine andamylase sedin theinvestigation © ExamTip Describing ancexplaining experimental result for enzyme experments's @eommon type of exam question so maesureyou understand whatis happening andcan felate this to changes inthe activesiteot the enzyme whenithas denatured, ofits low temperature, relate to the amount of kinetic energy themolecules have.Factors Affecting Enzyme Action: pH + The optimum pH formost enzymes is? © Some enzymes that are producediin acidic conditions, suchas the stomach, havea lower optimum pH (pH 2) © Somethatare produced inalkaline conditions, such as theduadenum, havea higher optimum pH (pH8 or) Ifthe pHs toohigh or toolow. the bonds that hold theamino acid chain togethertomake Uupthe protein can bedisrupted/destroyed + Thiswillchange the shape of the active site, so the substrate cannolonger fitintoit reducingtherate of activity ‘+ Moving t00 faraway from the optimum pH willcause the enzyme to denature andactivity wilstop SUBSTRATE Effect of pHon enzyme activityHeastosauemycsams coutormoresmesomeresources AT EXTREME gH IFURTHEST FROM OPTIMUM ENZYMES ARE COMPLETELY REMEMBER _THS DEPENDS. ON WHERE DENATURED THE ENZYME WORKS pees 2 2 MOVING AWAY FROM : eas : eee 3 oe ee : Een oy 2345678 90 R2B UH Bsavemyexams Graph showing the effect of pH on therate of activity foran enzyme from the duodenum © examTip Remember the terminology when witing about enzymes is very important, Make sure you refer to an enzyme becoming ‘denatured’ not ‘dying’ Being able to describe: AND explain theetfect of each envronmentalconditienan enzyme actions key Practise describing and explaining using the graphs and then check your descriptions against yournotes,Practical: Enzymes & pH + Amylaseis an enzyme that digests starch a polysacchaiide of glucose) intomaltose(a disaccharideof glucose) + The effect of citferent pH levels on the activity of amylase can be investigated Apparatus * Spotting tie + Measuring cylinder + TestTube + Syringe + Pipette + Stopwatch + Buffersolutions * Iodine + Starch solution . ‘Amylase solution Method + Adda cropof iodine toeach of thewellsofa spotting tle + Usea syringe to place 2m’ of amylaseinte a test tube + Add lem? of buffer solution at pH 2)tothe test tube usingasyringe + Usesnothertest tube to add 2.cm’ of starchsolution to the armylase and buffersolution, start the stopwatch whilst mixing using pipette + Every 10 seconds, transfera droplet of the solution toa new welloflodine solution (which should tum blue-black) + Repeat this transferprocess every 10 seconds unti the iodine solution stops tuning blue~ black (this means theamyiasehas broken down al the starch) + Record the time taken forthe reactiontobe completed + Repeat theinvestigation with buffers at different pH values (ranging from pH3.0 topH7.0)TNO REACTION maInvestigating the effect of pH onenzyme activity Results and Analysis + Amylaceis an enzymewhich breaks downstarch + When the iodine solution remains orange-brown, allthestarch has been digested + This investigation shows: © Atthe optimum pH, the iodine stoppeditumning blue-blackand remained orange- brownwithin the shortest amountof time. + This is because the enzymes working atts fastest rate andhas cicested all he starch + Athigher orlowerpH's (above orbslow the optimum) theiodine took a longartime tostop tumingbiue-biack or continued to tunblue-biack for theentireinvastigation © Thisis because on either side of the optimum pH, theenzymesaiesiaiting tobecome denatured and ae aresult are unable to bind with the starch orbreakitdown Limitations + Thestarch andamylasesolutions that need tobe used should be placedin a water bath at optimum temperaturebeforebeing used + Acolorimeter cenbe used tomeasure the progress of the reaction more accurately by measuring theabsorbanco/tranemission of ight through the colouredsolution © Acontrol ofiodine solution would be used forcomparison ‘AT EXTREME pH (FURTHEST FROM OPTIMUM) ENZYMES ARE COMPLETELY DENATURED RENEMEER TS DEPENDS ON WHERE THE ENZYME WORKS IN THE f0Dy ‘OPTIMUN MOVING AWAY FROM ‘OPTIMUM pit RESULTS. IN GRADUAL LOSS OF ACTIVITY AS ACTIVE SITE _BECONES DISTORTED FATE OF ENZYME ACTIVITY 123456738 902B% oH‘+ When working with practicalinvestigations, rememberto consider your CORMS evaluation WHAT IS BEING CHANGED IN THE INVESTIGATION (1.V) A CONTROL RELATED TO THE ORGANISM BEING USED (IF RELEVANT) REPEATS MUST BE CARRIED OUT FOR RELIABLE RESULTS HOW WILL YOU MEASURE YOUR DEPENDENT: VARIABLE (0.V/.) WHAT TIME SCALE WILL YOU USE (0.V.) WHAT WILL YOU CONTROL IN THE INVESTIGATION (C.V.) CORMSEvaluation + Inthis investigation, your evaluation should look something Ikethis: ° G-Weare changing thepH of the environment ° O-This's notrelevant to this investigationaswe aren't usingan organism ® R-Wewillrepeat the investigation several times toensure reliability > M1-Wewil measure thetimetaken for . » M2-theiodine to stop tuning black ® $-Wewill control the concentration and volume of the amylase, iodine and starch solutionusedin the investigation @ ExemTip When describing the effect of pHon enzyme activity, itis important to remember thatany pH outsideo the optimumean lead to the nzymebecomingpermanertly denatured.