diff --git a/.chloggen/featuregate_cli_python.yaml b/.chloggen/featuregate_cli_python.yaml
new file mode 100755
index 0000000000..3a444bd3ac
--- /dev/null
+++ b/.chloggen/featuregate_cli_python.yaml
@@ -0,0 +1,16 @@
+# One of 'breaking', 'deprecation', 'new_component', 'enhancement', 'bug_fix'
+change_type: breaking
+
+# The name of the component, or a single word describing the area of concern, (e.g. collector, target allocator, auto-instrumentation, opamp, github action)
+component: operator
+
+# A brief description of the change. Surround your text with quotes ("") if it needs to start with a backtick (`).
+note:  remove featuregate `operator.autoinstrumentation.python`. Use command line flag `--enable-python-instrumentation` instead
+
+# One or more tracking issues related to the change
+issues: [2582, 2672]
+
+# (Optional) One or more lines of additional information to render under the primary note.
+# These lines will be padded with 2 spaces and then inserted directly into the document.
+# Use pipe (|) for multiline entries.
+subtext:
diff --git a/internal/config/main.go b/internal/config/main.go
index e24e54c863..c8e24e8560 100644
--- a/internal/config/main.go
+++ b/internal/config/main.go
@@ -46,6 +46,7 @@ type Config struct {
 	enableMultiInstrumentation          bool
 	enableApacheHttpdInstrumentation    bool
 	enableDotNetInstrumentation         bool
+	enablePythonInstrumentation         bool
 	autoInstrumentationDotNetImage      string
 	autoInstrumentationGoImage          string
 	autoInstrumentationApacheHttpdImage string
@@ -82,6 +83,7 @@ func New(opts ...Option) Config {
 		enableMultiInstrumentation:          o.enableMultiInstrumentation,
 		enableApacheHttpdInstrumentation:    o.enableApacheHttpdInstrumentation,
 		enableDotNetInstrumentation:         o.enableDotNetInstrumentation,
+		enablePythonInstrumentation:         o.enablePythonInstrumentation,
 		targetAllocatorImage:                o.targetAllocatorImage,
 		operatorOpAMPBridgeImage:            o.operatorOpAMPBridgeImage,
 		targetAllocatorConfigMapEntry:       o.targetAllocatorConfigMapEntry,
@@ -132,6 +134,11 @@ func (c *Config) EnableDotNetAutoInstrumentation() bool {
 	return c.enableDotNetInstrumentation
 }
 
+// EnablePythonAutoInstrumentation is true when the operator supports dotnet auto instrumentation.
+func (c *Config) EnablePythonAutoInstrumentation() bool {
+	return c.enablePythonInstrumentation
+}
+
 // CollectorConfigMapEntry represents the configuration file name for the collector. Immutable.
 func (c *Config) CollectorConfigMapEntry() string {
 	return c.collectorConfigMapEntry
diff --git a/internal/config/options.go b/internal/config/options.go
index bfde188480..ab74059f04 100644
--- a/internal/config/options.go
+++ b/internal/config/options.go
@@ -45,6 +45,7 @@ type options struct {
 	enableMultiInstrumentation          bool
 	enableApacheHttpdInstrumentation    bool
 	enableDotNetInstrumentation         bool
+	enablePythonInstrumentation         bool
 	targetAllocatorConfigMapEntry       string
 	operatorOpAMPBridgeConfigMapEntry   string
 	targetAllocatorImage                string
@@ -99,6 +100,11 @@ func WithEnableDotNetInstrumentation(s bool) Option {
 		o.enableDotNetInstrumentation = s
 	}
 }
+func WithEnablePythonInstrumentation(s bool) Option {
+	return func(o *options) {
+		o.enablePythonInstrumentation = s
+	}
+}
 func WithTargetAllocatorConfigMapEntry(s string) Option {
 	return func(o *options) {
 		o.targetAllocatorConfigMapEntry = s
diff --git a/main.go b/main.go
index c52ca626a8..8042aaeed5 100644
--- a/main.go
+++ b/main.go
@@ -112,6 +112,7 @@ func main() {
 		enableMultiInstrumentation       bool
 		enableApacheHttpdInstrumentation bool
 		enableDotNetInstrumentation      bool
+		enablePythonInstrumentation      bool
 		collectorImage                   string
 		targetAllocatorImage             string
 		operatorOpAMPBridgeImage         string
@@ -138,6 +139,7 @@ func main() {
 	pflag.BoolVar(&enableMultiInstrumentation, "enable-multi-instrumentation", false, "Controls whether the operator supports multi instrumentation")
 	pflag.BoolVar(&enableApacheHttpdInstrumentation, constants.FlagApacheHttpd, true, "Controls whether the operator supports Apache HTTPD auto-instrumentation")
 	pflag.BoolVar(&enableDotNetInstrumentation, constants.FlagDotNet, true, "Controls whether the operator supports dotnet auto-instrumentation")
+	pflag.BoolVar(&enablePythonInstrumentation, constants.FlagPython, true, "Controls whether the operator supports python auto-instrumentation")
 	stringFlagOrEnv(&collectorImage, "collector-image", "RELATED_IMAGE_COLLECTOR", fmt.Sprintf("ghcr.io/open-telemetry/opentelemetry-collector-releases/opentelemetry-collector:%s", v.OpenTelemetryCollector), "The default OpenTelemetry collector image. This image is used when no image is specified in the CustomResource.")
 	stringFlagOrEnv(&targetAllocatorImage, "target-allocator-image", "RELATED_IMAGE_TARGET_ALLOCATOR", fmt.Sprintf("ghcr.io/open-telemetry/opentelemetry-operator/target-allocator:%s", v.TargetAllocator), "The default OpenTelemetry target allocator image. This image is used when no image is specified in the CustomResource.")
 	stringFlagOrEnv(&operatorOpAMPBridgeImage, "operator-opamp-bridge-image", "RELATED_IMAGE_OPERATOR_OPAMP_BRIDGE", fmt.Sprintf("ghcr.io/open-telemetry/opentelemetry-operator/operator-opamp-bridge:%s", v.OperatorOpAMPBridge), "The default OpenTelemetry Operator OpAMP Bridge image. This image is used when no image is specified in the CustomResource.")
@@ -180,6 +182,7 @@ func main() {
 		"enable-multi-instrumentation", enableMultiInstrumentation,
 		"enable-apache-httpd-instrumentation", enableApacheHttpdInstrumentation,
 		"enable-dotnet-instrumentation", enableDotNetInstrumentation,
+		"enable-python-instrumentation", enablePythonInstrumentation,
 	)
 
 	restConfig := ctrl.GetConfigOrDie()
@@ -199,6 +202,7 @@ func main() {
 		config.WithEnableMultiInstrumentation(enableMultiInstrumentation),
 		config.WithEnableApacheHttpdInstrumentation(enableApacheHttpdInstrumentation),
 		config.WithEnableDotNetInstrumentation(enableDotNetInstrumentation),
+		config.WithEnablePythonInstrumentation(enablePythonInstrumentation),
 		config.WithTargetAllocatorImage(targetAllocatorImage),
 		config.WithOperatorOpAMPBridgeImage(operatorOpAMPBridgeImage),
 		config.WithAutoInstrumentationJavaImage(autoInstrumentationJava),
diff --git a/pkg/constants/env.go b/pkg/constants/env.go
index 64efe59aa2..12391fde7e 100644
--- a/pkg/constants/env.go
+++ b/pkg/constants/env.go
@@ -37,4 +37,5 @@ const (
 
 	FlagApacheHttpd = "enable-apache-httpd-instrumentation"
 	FlagDotNet      = "enable-dotnet-instrumentation"
+	FlagPython      = "enable-python-instrumentation"
 )
diff --git a/pkg/featuregate/featuregate.go b/pkg/featuregate/featuregate.go
index d1b8057b6c..793c19ab1e 100644
--- a/pkg/featuregate/featuregate.go
+++ b/pkg/featuregate/featuregate.go
@@ -25,12 +25,6 @@ const (
 )
 
 var (
-	EnablePythonAutoInstrumentationSupport = featuregate.GlobalRegistry().MustRegister(
-		"operator.autoinstrumentation.python",
-		featuregate.StageBeta,
-		featuregate.WithRegisterDescription("controls whether the operator supports Python auto-instrumentation"),
-		featuregate.WithRegisterFromVersion("v0.76.1"),
-	)
 	EnableJavaAutoInstrumentationSupport = featuregate.GlobalRegistry().MustRegister(
 		"operator.autoinstrumentation.java",
 		featuregate.StageBeta,
diff --git a/pkg/instrumentation/podmutator.go b/pkg/instrumentation/podmutator.go
index 004e8239dd..0a4fbde291 100644
--- a/pkg/instrumentation/podmutator.go
+++ b/pkg/instrumentation/podmutator.go
@@ -253,7 +253,7 @@ func (pm *instPodMutator) Mutate(ctx context.Context, ns corev1.Namespace, pod c
 		logger.Error(err, "failed to select an OpenTelemetry Instrumentation instance for this pod")
 		return pod, err
 	}
-	if featuregate.EnablePythonAutoInstrumentationSupport.IsEnabled() || inst == nil {
+	if pm.config.EnablePythonAutoInstrumentation() || inst == nil {
 		insts.Python.Instrumentation = inst
 	} else {
 		logger.Error(nil, "support for Python auto instrumentation is not enabled")
diff --git a/pkg/instrumentation/podmutator_test.go b/pkg/instrumentation/podmutator_test.go
index 917eff5764..7343db7525 100644
--- a/pkg/instrumentation/podmutator_test.go
+++ b/pkg/instrumentation/podmutator_test.go
@@ -1266,6 +1266,7 @@ func TestMutatePod(t *testing.T) {
 					},
 				},
 			},
+			config: config.New(config.WithEnablePythonInstrumentation(true)),
 		},
 		{
 			name: "python injection multiple containers, true",
@@ -1533,6 +1534,7 @@ func TestMutatePod(t *testing.T) {
 					},
 				},
 			},
+			config: config.New(config.WithEnablePythonInstrumentation(true)),
 		},
 		{
 			name: "python injection feature gate disabled",
@@ -1619,13 +1621,6 @@ func TestMutatePod(t *testing.T) {
 					},
 				},
 			},
-			setFeatureGates: func(t *testing.T) {
-				originalVal := featuregate.EnablePythonAutoInstrumentationSupport.IsEnabled()
-				require.NoError(t, colfeaturegate.GlobalRegistry().Set(featuregate.EnablePythonAutoInstrumentationSupport.ID(), false))
-				t.Cleanup(func() {
-					require.NoError(t, colfeaturegate.GlobalRegistry().Set(featuregate.EnablePythonAutoInstrumentationSupport.ID(), originalVal))
-				})
-			},
 		},
 		{
 			name: "dotnet injection, true",
@@ -3844,6 +3839,7 @@ func TestMutatePod(t *testing.T) {
 			},
 			config: config.New(
 				config.WithEnableMultiInstrumentation(true),
+				config.WithEnablePythonInstrumentation(true),
 				config.WithEnableDotNetInstrumentation(true),
 			),
 		},
@@ -4506,6 +4502,7 @@ func TestMutatePod(t *testing.T) {
 			config: config.New(
 				config.WithEnableMultiInstrumentation(true),
 				config.WithEnableDotNetInstrumentation(true),
+				config.WithEnablePythonInstrumentation(true),
 			),
 		},
 		{
diff --git a/pkg/instrumentation/upgrade/upgrade.go b/pkg/instrumentation/upgrade/upgrade.go
index 831271c2a8..ae0cd1789c 100644
--- a/pkg/instrumentation/upgrade/upgrade.go
+++ b/pkg/instrumentation/upgrade/upgrade.go
@@ -34,7 +34,6 @@ var (
 	defaultAnnotationToGate = map[string]*featuregate2.Gate{
 		constants.AnnotationDefaultAutoInstrumentationJava:   featuregate.EnableJavaAutoInstrumentationSupport,
 		constants.AnnotationDefaultAutoInstrumentationNodeJS: featuregate.EnableNodeJSAutoInstrumentationSupport,
-		constants.AnnotationDefaultAutoInstrumentationPython: featuregate.EnablePythonAutoInstrumentationSupport,
 		constants.AnnotationDefaultAutoInstrumentationGo:     featuregate.EnableGoAutoInstrumentationSupport,
 		constants.AnnotationDefaultAutoInstrumentationNginx:  featuregate.EnableNginxAutoInstrumentationSupport,
 	}
@@ -63,6 +62,7 @@ func NewInstrumentationUpgrade(client client.Client, logger logr.Logger, recorde
 	defaultAnnotationToConfig := map[string]autoInstConfig{
 		constants.AnnotationDefaultAutoInstrumentationApacheHttpd: {constants.FlagApacheHttpd, cfg.EnableApacheHttpdAutoInstrumentation()},
 		constants.AnnotationDefaultAutoInstrumentationDotNet:      {constants.FlagDotNet, cfg.EnableDotNetAutoInstrumentation()},
+		constants.AnnotationDefaultAutoInstrumentationPython:      {constants.FlagPython, cfg.EnablePythonAutoInstrumentation()},
 	}
 
 	return &InstrumentationUpgrade{
@@ -132,6 +132,11 @@ func (u *InstrumentationUpgrade) upgrade(_ context.Context, inst v1alpha1.Instru
 						upgraded.Spec.DotNet.Image = u.DefaultAutoInstDotNet
 						upgraded.Annotations[annotation] = u.DefaultAutoInstDotNet
 					}
+				case constants.AnnotationDefaultAutoInstrumentationPython:
+					if inst.Spec.Python.Image == autoInst {
+						upgraded.Spec.Python.Image = u.DefaultAutoInstPython
+						upgraded.Annotations[annotation] = u.DefaultAutoInstPython
+					}
 				}
 			} else {
 				u.Logger.Error(nil, "autoinstrumentation not enabled for this language", "flag", config.id)
diff --git a/pkg/instrumentation/upgrade/upgrade_test.go b/pkg/instrumentation/upgrade/upgrade_test.go
index 9e9aa7547f..546c7d3e40 100644
--- a/pkg/instrumentation/upgrade/upgrade_test.go
+++ b/pkg/instrumentation/upgrade/upgrade_test.go
@@ -80,6 +80,7 @@ func TestUpgrade(t *testing.T) {
 			config.WithAutoInstrumentationNginxImage("nginx:1"),
 			config.WithEnableApacheHttpdInstrumentation(true),
 			config.WithEnableDotNetInstrumentation(true),
+			config.WithEnablePythonInstrumentation(true),
 		),
 	).Default(context.Background(), inst)
 	assert.Nil(t, err)
@@ -103,6 +104,7 @@ func TestUpgrade(t *testing.T) {
 		config.WithAutoInstrumentationNginxImage("nginx:2"),
 		config.WithEnableApacheHttpdInstrumentation(true),
 		config.WithEnableDotNetInstrumentation(true),
+		config.WithEnablePythonInstrumentation(true),
 	)
 	up := NewInstrumentationUpgrade(k8sClient, ctrl.Log.WithName("instrumentation-upgrade"), &record.FakeRecorder{}, cfg)